Genetic Engineering Publications - GEG Tech top picks
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December 19, 2014 1:22 AM
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7q11.23 dosage-dependent dysregulation in human pluripotent stem cells affects transcriptional programs in disease-relevant lineages - Nature Genetics

7q11.23 dosage-dependent dysregulation in human pluripotent stem cells affects transcriptional programs in disease-relevant lineages - Nature Genetics | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Giuseppe Testa and colleagues report the generation and transcriptional characterization of patient-derived induced pluripotent stem cells (iPSCs) with copy number variants at 7q11.23, which cause syndromes including neurocognitive phenotypes. They find that the dosage of the transcription factor gene GTF2I accounts for 10-20% of the transcriptional dysregulation observed in these cells.
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November 7, 6:46 AM
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Innovative immunotherapy harnesses natural killer T cells to combat solid tumors

Innovative immunotherapy harnesses natural killer T cells to combat solid tumors | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
In the fight against cancer, chimeric antigen receptor T cell (CAR-T) therapy has achieved notable success in treating blood cancers. However, it has been largely ineffective against solid tumors.
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In the fight against cancer, CAR-T cells have achieved remarkable results in treating blood cancers. However, they have proven largely ineffective against solid tumors. CAR-NKT cells, on the other hand, possess innate properties that make them particularly well-equipped to combat solid tumors. Specifically, they express a T cell receptor that recognizes glycolipid antigens presented by CD1d molecules. CAR-NKT cells effectively eliminate CD1d-expressing M2 macrophages in the tumor microenvironment. These macrophages promote tumor growth and suppress immune responses, making their elimination essential for strengthening antitumor immunity. By targeting these cells, CAR-NKT therapy effectively reprograms the tumor microenvironment from a pro-tumor to an anti-tumor state. CAR-NKT cells promote epitope spreading, a process by which the immune system recognizes and attacks new targets, activating T-cell responses. Researchers further demonstrated that combining CAR-NKT cells with PD1 blockade significantly enhances their antitumor activity, and that this activity can be further strengthened by vaccination approaches, such as those using dendritic cells loaded with alpha-galactosylceramide. 

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October 28, 11:13 AM
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RNA Repeat Targeting as Potential New Therapeutic Strategy for ALS and Frontotemporal Dementia

RNA Repeat Targeting as Potential New Therapeutic Strategy for ALS and Frontotemporal Dementia | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Researchers in the US report a potentially new therapeutic strategy for amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Using an optimised Cas13d-based RNA targeting system, they demonstrate significant in vitro and in vivo reduction of the RNA repeat mutations most commonly identified in patients with ALS or FTD.
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The hexanucleotide GGGGCC repeat expansion in the C9orf72 gene is the most common identified mutation in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) patients. This expansion leads to the production of repeat RNA and dipeptide repeat proteins (DPR), both of which contribute to disease and are recognised as important disease markers. In a recent study, a team in the United States demonstrated that CRISPR-Cas13d can be optimised to specifically target GGGGCC repeat RNA to minimise the translation of associated poly-dipeptides.The team shows that the optimised CRISPR-Cas13d system effectively reduced DPR production when delivered via lentiviral particles to various in vitro models, including C9orf72-ALS patient-derived induced pluripotent stem cells (iPSCs) and motor neurons derived from those iPSCs, and that the efficiency of RNA targeting corresponded to Cas13d expression levels. Intracerebroventricular injection of the C9orf72-targeting cargo (delivered via AAV9) in C9orf72 repeat transgenic mice revealed a significant reduction in DPR levels. The team reports that the approach exhibits specificity for expanded repeats, with minimal effect on wild-type C9orf72 transcripts and protein levels.

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October 8, 9:48 AM
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World-first therapy using donor cells sends autoimmune diseases into remission

World-first therapy using donor cells sends autoimmune diseases into remission | Genetic Engineering Publications - GEG Tech top picks | Scoop.it

The treatment’s success in three people raises hopes for mass production of cutting-edge CAR T therapies.

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The treatment’s success in three people raises hopes for mass production of cutting-edge CAR T therapies.

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September 30, 6:02 AM
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New discovery offers hope for overcoming CAR-T cell therapy resistance

New discovery offers hope for overcoming CAR-T cell therapy resistance | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Mayo Clinic researchers mined the molecular foundations of cancer and uncovered a new reason chimeric antigen receptor (CAR-T cell therapy) fails in some patients.
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CAR-T cell therapy works very well in only about a third of cancer patients. One of the main factors in failure is T-cell depletion. This condition causes many patients to relapse within a year of CAR-T cell therapy. In search of new solutions, a research team compared data from patients in remission with those whose CAR-T cell therapy had failed. They also studied how CAR-T cells killed tumors grown in laboratory mice. They compared the results of mice that responded well to CAR-T therapy with those that did not. The team observed an increase in interleukin-4 (IL-4) protein in both human and mouse CAR-T cell depletion samples. The team then used CRISPR gene-editing technology to eliminate or modify the IL-4 protein causing CAR-T cell dysfunction. The researchers also tested monoclonal antibodies to block or neutralize the IL-4 protein. They found that they also rejuvenated CAR-T cells and their ability to block cancer. 

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September 10, 5:06 AM
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CD5 knockout using CRISPR boosts CAR T cell therapy efficacy

CD5 knockout using CRISPR boosts CAR T cell therapy efficacy | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The effectiveness of CAR T cell therapy against a variety of cancers, including solid tumors, could be boosted greatly by using CRISPR-Cas9 technology to knock out the gene for CD5, a protein found on the surface of T cells, according to a preclinical study from investigators at the University of Pennsylvania's Perelman School of Medicine and Abramson Cancer Center.
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CAR T cells are T cells designed to attack specific targets on cancer cells. They have produced remarkable results in certain patients with blood cancers. However, they have not worked well against other cancers, particularly solid tumor cancers such as pancreatic cancer, prostate cancer, and melanoma. The researchers looked for techniques to increase the effectiveness of CAR T cell therapy. The study, published today in Science Immunology, suggests that using CRISPR-Cas9 technology to knock out the CD5 gene could be a first-rate technique. By shedding light on the hitherto obscure role of the CD5 protein present on the surface of T cells, the researchers discovered that it functions as a robust immune checkpoint, limiting the efficiency of T lymphocytes. By deleting it, they showed that the anti-cancer activity of CAR T cells improved considerably in various preclinical cancer models. A Phase I clinical trial of CD5-knockout CAR T cells will soon begin enrolling patients with CD5-bearing T-cell lymphoma.

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September 6, 3:50 AM
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Cellular immunotherapy offers hope for solid tumors

Cellular immunotherapy offers hope for solid tumors | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Early findings from a small clinical trial provide evidence that a new cellular immunotherapy approach may be effective in treating metastatic solid tumors. In the trial, researchers from the National Institutes of Health (NIH) genetically engineered normal white blood cells, known as lymphocytes, from each patient to produce receptors that recognize and attack their specific cancer cells.
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To date, no effective cell therapy for solid cancers has been found. However, initial results from a small clinical trial show that a new cellular immunotherapy approach could effectively treat metastatic solid tumors. Researchers collected lymphocytes from each patient's tumor as part of the trial. They then used sophisticated molecular characterization techniques to identify and isolate the receptors on these lymphocytes, called T-cell receptors, which recognized specific changes in each patient's tumor. After genetically sequencing these receptors, they used a retrovirus to insert the receptor genes into normal lymphocytes taken from each patient's circulating blood. The initial results came from people with metastatic colorectal cancer who had already undergone several previous treatments. Personalized immunotherapy shrank tumors in several patients and prevented tumors from growing back for up to seven months. The results were published on 11 July 2024 in Nature Medicine.

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July 11, 9:12 AM
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In situ targeted base editing of bacteria in the mouse gut - Nature

In situ targeted base editing of bacteria in the mouse gut - Nature | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Microbiome research is now demonstrating a growing number of bacterial strains and genes that affect our health1. Although CRISPR-derived tools have shown great success in editing disease-driving genes in human cells2, we currently lack the tools to achieve comparable success for bacterial targets in situ. Here we engineer a phage-derived particle to deliver a base editor and modify Escherichia coli colonizing the mouse gut. Editing of a β-lactamase gene in a model E. coli strain resulted in a median editing efficiency of 93% of the target bacterial population with a single dose. Edited bacteria were stably maintained in the mouse gut for at least 42 days following treatment. This was achieved using a non-replicative DNA vector, preventing maintenance and dissemination of the payload. We then leveraged this approach to edit several genes of therapeutic relevance in E. coli and Klebsiella pneumoniae strains in vitro and demonstrate in situ editing of a gene involved in the production of curli in a pathogenic E. coli strain. Our work demonstrates the feasibility of modifying bacteria directly in the gut, offering a new avenue to investigate the function of bacterial genes and opening the door to the design of new microbiome-targeted therapies. Edited bacteria were stably maintained in mouse gut for at least 42 days following the delivery of a base editor using an engineered phage-derived particle to modify Escherichia coli colonizing the gut.
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A group from Paris-based Eligo Bioscience has engineered a phage-derived vector to deliver a base editor and modify Escherichia coli while they are colonizing the mouse gut. It marks the first time the genomes of bacteria have been base edited, precisely and efficiently, directly in the gut. 

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June 17, 5:05 AM
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RNA editing using CRISPRs shows promise for genetic disease treatment

RNA editing using CRISPRs shows promise for genetic disease treatment | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
A team at Montana State University published research this week that shows how RNA, the close chemical cousin to DNA, can be edited using CRISPRs.
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RNA repair could be a fundamental aspect of biology, and harnessing this activity could lead to new life-saving cures. In particular, RNA editing has important applications in the search for treatments for genetic diseases. A team of researchers recently published research showing how RNA can be edited using a different CRISPR system to that for DNA, called type III. The study article entitled "Repair of CRISPR-guided RNA breaks enables site-specific RNA excision in human cells" was published in the journal Science and is the latest advance in the team's ongoing exploration of CRISPR applications for programmable genetic engineering. 

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April 24, 5:41 AM
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How to supercharge cancer-fighting cells: give them stem-cell skills - Nature

How to supercharge cancer-fighting cells: give them stem-cell skills - Nature | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The bioengineered immune players called CAR T cells last longer and work better if pumped up with a large dose of a protein that makes them resemble stem cells.
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Keeping cells active long enough to eliminate cancer has proved difficult, particularly in solid tumors such as those of the breast and lung. Scientists are therefore looking for better ways to help CAR T cells multiply faster and last longer in the body. To this end, researchers compared samples of CAR T cells used to treat people with leukemia. They analyzed the role of cellular proteins that regulate gene activity and serve as master switches in T cells. They discovered a set of 41 genes that were more active in CAR T cells associated with a good response to treatment than in cells associated with a poor response. All 41 genes appeared to be regulated by a master protein called FOXO1. The researchers then engineered the CAR T cells to produce more FOXO1 than usual. Gene activity in these cells began to resemble that of memory T stem cells, which recognize cancer and respond quickly to it. The researchers then injected the modified cells into mice with different types of cancer. Extra FOXO1 enabled the CAR T cells to better reduce both solid tumors and blood cancers. Moreover, another team also reached the same conclusion by working on gene activity analysis in CAR T cells and also discovered that IL-15 activated genes associated with FOXO1. 

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April 12, 6:58 AM
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New genetic system analyzes the underlying mechanisms of CRISPR-based DNA repair outcomes

New genetic system analyzes the underlying mechanisms of CRISPR-based DNA repair outcomes | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Since its breakthrough development more than a decade ago, CRISPR has revolutionized DNA editing across a broad range of fields.
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Researchers have developed a new genetic system to test and analyze the mechanisms underlying the results of CRISPR-based DNA repair. As described in Nature Communications, they have developed a sequence analyzer to help track on- and off-target mutational changes and how they are inherited from one generation to the next. The tool, called Integrated Classifier Pipeline (ICP), can reveal specific categories of mutations resulting from CRISPR editing. Developed in flies and mosquitoes, ICP provides a "fingerprint" of how genetic material is inherited, enabling scientists to track the source of mutational changes and the associated risks emerging from potentially problematic modifications. PKI can help unravel the complex biological issues that arise when determining the mechanisms behind CRISPR. Although developed in insects, ICP has great potential for human applications.

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MRI and lumbar puncture may not be necessary for managing CAR T-cell associated toxicities

MRI and lumbar puncture may not be necessary for managing CAR T-cell associated toxicities | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Magnetic resonance imaging (MRI) and lumbar puncture (LP) may not always be necessary for diagnosing and managing a serious neurological complication associated with CAR T-cell therapy, according to a new Blood Advances study.
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A study published in Blood Advances reveals that magnetic resonance imaging (MRI) and lumbar puncture (LP) may not be systematically necessary in the diagnosis and management of severe cases of neurotoxicity linked to CAR T cell therapy. Instead, electroencephalogram (EEG), a non-invasive test, has proved useful in the management of these complications. The researchers examined the usefulness of these tests in 190 patients treated with CAR-T at Rennes University Hospital, where during treatment around 48% of patients developed immune effector cell-associated neurotoxicity syndrome (ICANS). The researchers assessed how the different tests affected patient treatment, such as how medications, e.g. antibiotics and anti-epileptic treatments, were prescribed based on abnormal results, and how these treatments altered patient outcomes. The results ultimately revealed that abnormal findings were more common in patients with more severe ICANS. MRI findings were often normal, and although LP and EEG often showed abnormalities, they were more common in more severe cases of ICANS. When it came to therapeutic decisions, MRI rarely led to changes, LP sometimes led to unnecessary treatments in cases of suspected infections, and EEG often resulted in adjustments to antiepileptic drugs.

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March 21, 6:48 AM
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News: CRISPR Screen Unlocks Cancer Metastasis Mystery

News: CRISPR Screen Unlocks Cancer Metastasis Mystery | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
In vivo CRISPR genome-wide screening pinpoints the transcriptional modulator CITED2 as a pivotal driver in the progression of prostate cancer to bone metastasis.
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In vivo screening of the CRISPR genome identifies the transcriptional modulator CITED2 as an essential factor in the progression of prostate cancer to bone metastases. The discovery not only improves understanding of the molecular basis of the disease, but also opens up new avenues for targeted therapies, potentially revolutionizing treatment paradigms for patients battling advanced prostate cancer. The study meticulously engineered non-metastatic human prostate cancer cell lines to activate or inhibit gene expression using CRISPRa or CRISPRi technology. Modified cancer cells were then implanted into the prostate of nude mice, and following tumor development and emergence of metastases, primary and metastatic tumors were harvested for analysis. In vivo CRISPR screening identified CITED2 as an important promoter of bone metastasis, standing out among various genes for its substantial impact. Subsequent functional validation experiments, including innovative organ-on-a-chip assays, reinforced CITED2's role in promoting bone invasion, highlighting its potential as a therapeutic target. The research also looked at CITED2-driven transcriptional profiles, revealing distinct patterns of primary and metastatic cancer, which could inform the development of precision medicine approaches. 

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March 11, 6:11 AM
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Breakthrough CRISPR technique unlocks insights about cancer immunology

Breakthrough CRISPR technique unlocks insights about cancer immunology | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Over the past two decades, the immune system has attracted increasing attention for its role in fighting cancer.
BigField GEG Tech's insight:

CRISPR-based gene editing has become a mainstay of biological discovery, providing relatively rapid insight into the function of individual genes and targets for new therapies. However, the main challenge is that it is difficult to edit immune cells without changing their biology, hampering the ability to study immune cell behavior in all its complexity in a living organism. In an earlier study, researchers used CHimeric IMmune Editing (CHIME) to knock out a gene called Ptpn2, which has shown promise for cancer immunotherapy. The recent study published in Nature Immunology explores methods to increase the precision and versatility of CHIME, including simultaneous deletion of multiple genes and targeted application in specific cell types, also using CRISPR to disrupt genes in the edited cells once they were already back inside the animal. This research has successfully demonstrated the feasibility of various innovative approaches to genetic manipulation, offering new avenues for the study of immune gene function. 

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Scooped by BigField GEG Tech
October 30, 7:29 AM
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Buoyed by ‘milestone’ clinical result, RNA editing is poised to treat diseases | Science | AAAS

Buoyed by ‘milestone’ clinical result, RNA editing is poised to treat diseases | Science | AAAS | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Multiple approaches aim to counteract mutant genes by fixing the faulty RNA they produce
BigField GEG Tech's insight:

The field of RNA editing may be in its infancy, but pharma companies are already testing its use in some types of eye disease and cancer.

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October 23, 9:30 AM
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Innovative CAR-T cells effectively target glioblastoma with reduced toxicity

Innovative CAR-T cells effectively target glioblastoma with reduced toxicity | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Glioblastoma is the most common and most aggressive primary brain tumor, with an average survival after diagnosis of less than two years, and against which current treatments remain ineffective.
BigField GEG Tech's insight:

Glioblastoma is the most common and aggressive primary brain tumor, with an average survival after diagnosis of less than two years, and against which current treatments remain ineffective. In recent years, immunotherapies have given patients hope, albeit with relatively modest success. One research team has succeeded in identifying a specific marker, PTPRZ1, on the surface of tumour cells, and generating CAR-T cells to destroy them. Moreover, this CAR-T cell-based therapy appears to be able to target diseased tumor cells that do not carry this specific marker, while sparing healthy cells. The team tested the treatment in vivo on mouse models of human glioblastoma. Tumor growth was controlled, remarkably prolonging the life of the mice without signs of toxicity. These results, published in the journal Cancer Immunology Research, represent a first step towards the development of clinical trials with human patients. 

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October 4, 5:22 AM
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CRISPR helps brain stem cells regain youth in mice

CRISPR helps brain stem cells regain youth in mice | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Disabling a gene involved in metabolism rejuvenates cells’ ability to spin off new neurons.
BigField GEG Tech's insight:

Clues to keeping the brain’s regenerative cells youthful and energetic into old age have emerged by applying CRISPR gene editing to mice

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Scooped by BigField GEG Tech
September 23, 8:16 AM
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Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells | Nature Medicine

Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells | Nature Medicine | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Resistance to genotoxic therapies and tumor recurrence are hallmarks of glioblastoma (GBM), an aggressive brain tumor. In this study, we investigated functional drivers of post-treatment recurrent GBM through integrative genomic analyses, genome-wide genetic perturbation screens in patient-derived GBM models and independent lines of validation. Specific genetic dependencies were found consistent across recurrent tumor models, accompanied by increased mutational burden and differential transcript and protein expression compared to its primary GBM predecessor. Our observations suggest a multi-layered genetic response to drive tumor recurrence and implicate PTP4A2 (protein tyrosine phosphatase 4A2) as a modulator of self-renewal, proliferation and tumorigenicity in recurrent GBM. Genetic perturbation or small-molecule inhibition of PTP4A2 acts through a dephosphorylation axis with roundabout guidance receptor 1 (ROBO1) and its downstream molecular players, exploiting a functional dependency on ROBO signaling. Because a pan-PTP4A inhibitor was limited by poor penetrance across the blood–brain barrier in vivo, we engineered a second-generation chimeric antigen receptor (CAR) T cell therapy against ROBO1, a cell surface receptor enriched across recurrent GBM specimens. A single dose of ROBO1-targeted CAR T cells doubled median survival in cell-line-derived xenograft (CDX) models of recurrent GBM. Moreover, in CDX models of adult lung-to-brain metastases and pediatric relapsed medulloblastoma, ROBO1 CAR T cells eradicated tumors in 50–100% of mice. Our study identifies a promising multi-targetable PTP4A–ROBO1 signaling axis that drives tumorigenicity in recurrent GBM, with potential in other malignant brain tumors. Functional CRISPR screens in patient-matched pre-treatment and post-treatment glioblastoma models identify the PTP4A–ROBO1 axis as a driver of tumorigenicity and enriched ROBO1 expression in recurrent glioblastoma that can be targeted with CAR T cell therapy.
BigField GEG Tech's insight:

With existing treatments such as surgery, radiotherapy, and chemotherapy, brain tumors often recur, and patients' survival is limited to just a few months. However, a team of researchers has discovered a new pathway cancer cells use to infiltrate the brain. To find this pathway, the researchers used large-scale gene-editing technology to compare genetic dependencies, particularly in glioblastoma at initial diagnosis and after its recurrence following standard treatments. This new pathway is used for axonal guidance, a signaling axis that helps establish standard brain architecture, which can be invaded by cancer cells. Their research also reveals that blocking this pathway using CAR-T cells, mainly targeting the Roundabout Guidance Receptor 1 protein, looks promising for stopping and killing these tumors. The treatment was tested in mouse models on three different types of cancer: adult glioblastoma, adult lung-cerebral metastases, and pediatric medulloblastoma. In all three models, the treatment doubled survival time. In two of the three diseases, the tumor was eradicated in at least 50% of the mice. 

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September 9, 5:17 AM
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CD22-targeted CAR-T therapy shows promising results in relapsed large B-cell lymphoma

CD22-targeted CAR-T therapy shows promising results in relapsed large B-cell lymphoma | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
CAR-T cell therapy, which targets a specific protein on the surface of cancer cells, causes tumors to shrink or disappear in about half of patients with large B-cell lymphoma who haven't experienced improvement with chemotherapy treatments.
BigField GEG Tech's insight:

One of CAR-T cell therapy targets CD19 on the surface of lymphoma cells. It causes tumors to shrink or disappear in around half of patients with large B-cell lymphoma who have not improved with chemotherapy treatments. If this CAR-T treatment fails or the cancer recurs, as it does in around half of patients, the prognosis is dire. The median survival time after a relapse is around six months. However, a phase 1 clinical trial conducted at Stanford Medicine showed that a new CAR-T cell therapy targeting CD22 on the surface of cancer cells significantly improved outcomes for these patients: more than half of the 38 people enrolled in the trial, 37 of whom had already relapsed after initial CAR-T therapy, experienced a complete response to their cancer. Over half of the patients treated lived for at least two years after treatment.

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July 31, 9:08 AM
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Brainwide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic editor

Brainwide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic editor | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Prion disease is caused by misfolding of the prion protein (PrP) into pathogenic self-propagating conformations, leading to rapid-onset dementia and death. However, elimination of endogenous PrP halts prion disease progression. In this study, we describ
BigField GEG Tech's insight:

Prion disease, which leads to rapid neurodegeneration and death, is caused by the presence of deformed versions of the prion protein. These cause a cascade effect in the brain: the defective prion proteins deform other proteins, and together these proteins not only cease to function correctly but also form toxic aggregates that kill neurons—most conventional drugs work by targeting just one protein. However, in less than two years, using a research tool called CRISPRoff, researchers have developed a set of molecular tools called CHARMs that act upstream, disabling the gene that codes for a defective protein so that the protein is never produced. CHARMs achieve this by epigenetic editing. Unlike gene editing, epigenetic editing does not alter the underlying DNA, and the gene itself remains intact. Furthermore, like gene editing, epigenetic editing is stable, meaning that a gene switched off by CHARM should remain switched off. This would mean that patients would only have to take CHARM once, unlike other drugs. Moreover, to address the transmission problem, the researchers have also designed an AAV vector that can enter the brain more efficiently by exploiting a pathway that naturally transports iron in the brain.  

 

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June 27, 5:54 AM
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Bridge RNAs direct programmable recombination of target and donor DNA - Nature

Bridge RNAs direct programmable recombination of target and donor DNA - Nature | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Genomic rearrangements, encompassing mutational changes in the genome such as insertions, deletions or inversions, are essential for genetic diversity. These rearrangements are typically orchestrated by enzymes that are involved in fundamental DNA repair processes, such as homologous recombination, or in the transposition of foreign genetic material by viruses and mobile genetic elements1,2. Here we report that IS110 insertion sequences, a family of minimal and autonomous mobile genetic elements, express a structured non-coding RNA that binds specifically to their encoded recombinase. This bridge RNA contains two internal loops encoding nucleotide stretches that base-pair with the target DNA and the donor DNA, which is the IS110 element itself. We demonstrate that the target-binding and donor-binding loops can be independently reprogrammed to direct sequence-specific recombination between two DNA molecules. This modularity enables the insertion of DNA into genomic target sites, as well as programmable DNA excision and inversion. The IS110 bridge recombination system expands the diversity of nucleic-acid-guided systems beyond CRISPR and RNA interference, offering a unified mechanism for the three fundamental DNA rearrangements—insertion, excision and inversion—that are required for genome design. A bispecific non-coding RNA expressed by the IS110 family of mobile genetic elements forms the basis of a programmable genome-editing system that enables the insertion, excision or inversion of specific target DNA sequences.
BigField GEG Tech's insight:

RNA-guided recombinase enzymes have been discovered that herald a new chapter for genome editing — enabling the insertion, inversion or deletion of long DNA sequences at user-specified genome positions. 

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Scooped by BigField GEG Tech
June 17, 4:43 AM
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RNA editing using CRISPRs shows promise for genetic disease treatment

RNA editing using CRISPRs shows promise for genetic disease treatment | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
A team at Montana State University published research this week that shows how RNA, the close chemical cousin to DNA, can be edited using CRISPRs.
BigField GEG Tech's insight:

RNA repair could be a fundamental aspect of biology, and harnessing this activity could lead to new life-saving cures. In particular, RNA editing has important applications in the search for treatments for genetic diseases. A team of researchers recently published research showing how RNA can be edited using a different CRISPR system to that for DNA, called type III. The study article entitled "Repair of CRISPR-guided RNA breaks enables site-specific RNA excision in human cells" was published in the journal Science and is the latest advance in the team's ongoing exploration of CRISPR applications for programmable genetic engineering. 

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Scooped by BigField GEG Tech
April 15, 9:55 AM
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New immunotherapy strategy potentially lays the groundwork for treating autoimmune diseases

New immunotherapy strategy potentially lays the groundwork for treating autoimmune diseases | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Mayo Clinic scientists have developed an immunotherapy strategy that potentially lays the groundwork for treating a spectrum of autoimmune diseases.
BigField GEG Tech's insight:

Graft-versus-host disease occurs when a donor's cells attack the recipient's tissues, usually following a bone marrow or stem cell transplant. In a recent study, a new technique involving the combination of Chimeric Antigen Receptors (CARs) with Mesenchymal Stromal Cells (MSCs), resulting in modified stem cells known as CAR-MSCs, was used to specifically target a protein linked to graft-versus-host disease, but also to inflammatory bowel diseases such as ulcerative colitis and Crohn's disease. In mouse models, when stimulated by the specific protein for which they were designed, CAR-MSCs showed an enhanced ability to reach the inflamed area, better control inflammation and improve outcome and survival. This was mediated by a change in the genetic signature of CAR-MSCs, the proteins they released and receptor expression. These preliminary results pave the way for future applications of this technology, paving the way for improving the versatility of the therapy to treat various diseases across the autoimmune spectrum.

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Scooped by BigField GEG Tech
April 11, 9:22 AM
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Novel CAR T-Cell Therapy Displays Activity in Glioblastoma

Novel CAR T-Cell Therapy Displays Activity in Glioblastoma | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Treatment with a next-generation CAR T-cell agent displayed early efficacy in a small group of patients with glioblastoma.
BigField GEG Tech's insight:

CARv3-TEAM-E T cells are CAR T cells targeting EGFR variant III tumor-specific antigen (EGFRvIII) in addition to wild-type EGFR. According to the results of the phase 1 INCIPIENT trial (NCT05660369) published in the New England Journal of Medicine, preliminary results in humans demonstrated that all 3 glioblastoma patients treated with CARv3-TEAM-E T cells between March 2023 and July 2023 showed dramatic and rapid radiographic regression of their tumors within days of receiving CARv3-TEAM-E T cells via a single intraventricular infusion. Responses were transient in 2 of the patients, however 1 patient, a 72-year-old man, showed an 18.5% decrease in tumor cross-sectional area on day 2 after receiving a single infusion of 10 x 106 CAR-positive CARv3-TEAM-E T cells. Moreover, 69 days after infusion, tumor cross-sectional area had decreased by a further 60.7% from baseline; the response was sustained and continued to improve at the last assessment, which took place more than 150 days after infusion. 

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Scooped by BigField GEG Tech
March 29, 7:05 AM
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Novel dual-target CAR T cell therapy shows promise in treating recurrent glioblastoma

Novel dual-target CAR T cell therapy shows promise in treating recurrent glioblastoma | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Targeting two brain tumor-associated proteins-;rather than one-;with CAR T cell therapy shows promise as a strategy for reducing solid tumor growth in patients with recurrent glioblastoma (GBM), an aggressive form of brain cancer, according to early results from the first six patients treated in an ongoing Phase I clinical trial led by researchers from the Perelman School of Medicine at the University of Pennsylvania and Penn Medicine's Abramson Cancer Center.
BigField GEG Tech's insight:

Glioblastoma (GBM) is the most common and aggressive type of cancerous brain tumor in adults. People with GBM generally expect to live 12 to 18 months after diagnosis. Despite decades of research, there is no known cure for GBM, and treatments have only a limited effect on extending an individual's life expectancy. However, researchers have tested a technology that delivers CAR-T cells targeting two proteins commonly found in brain tumors: epidermal growth factor receptor (EGFR), estimated to be present in 60% of all GBMs, and interleukin-13 receptor alpha 2 (IL13Rα2), which is expressed in over 75% of GBMs. While CAR-T cell therapy for blood cancers is usually administered intravenously, the researchers administered these dual-targeted CAR-T cells intrathecally, by injection into the cerebrospinal fluid, so that the modified cells could reach the tumors more directly in the brain. Magnetic Resonance Imaging scans taken 24 to 48 hours after administration of dual-targeted CAR-T cells targeting EGFR and IL13Rα2 revealed a reduction in tumor size in all six patients, and these reductions were maintained up to several months later in a subgroup of patients. 

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Scooped by BigField GEG Tech
March 13, 8:21 AM
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A non-FRET DNA reporter that changes fluorescence colour upon nuclease digestion | Nature Nanotechnology

A non-FRET DNA reporter that changes fluorescence colour upon nuclease digestion | Nature Nanotechnology | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Fluorescence resonance energy transfer (FRET) reporters are commonly used in the final stages of nucleic acid amplification tests to indicate the presence of nucleic acid targets, where fluorescence is restored by nucleases that cleave the FRET reporters. However, the need for dual labelling and purification during manufacturing contributes to the high cost of FRET reporters. Here we demonstrate a low-cost silver nanocluster reporter that does not rely on FRET as the on/off switching mechanism, but rather on a cluster transformation process that leads to fluorescence color change upon nuclease digestion. Notably, a 90 nm red shift in emission is observed upon reporter cleavage, a result unattainable by a simple donor-quencher FRET reporter. Electrospray ionization–mass spectrometry results suggest that the stoichiometric change of the silver nanoclusters from Ag13 (in the intact DNA host) to Ag10 (in the fragments) is probably responsible for the emission colour change observed after reporter digestion. Our results demonstrate that DNA-templated silver nanocluster probes can be versatile reporters for detecting nuclease activities and provide insights into the interactions between nucleases and metallo-DNA nanomaterials. Here the authors present a non-FRET DNA-templated silver nanocluster probe that exhibits a distinct colour switch from green to red upon nuclease digestion, visible under UV excitation, offering a low-cost, effective alternative to fluorescent reporters for detecting nuclease activities.
BigField GEG Tech's insight:

A new tool could reduce the cost of diagnosing infectious diseases. Researchers have developed a new, less expensive means of detecting nuclease digestion, one of the critical steps in many nucleic acid detection applications, such as those used to identify COVID-19 and other infectious diseases. A new study published in the journal Nature Nanotechnology shows that this inexpensive tool, called Subak, is effective in determining when nucleic acid cleavage occurs, which happens when an enzyme called nuclease breaks down nucleic acids, such as DNA or RNA, into smaller fragments. The traditional method for identifying nuclease activity, the Fluorescence Resonance Energy Transfer (FRET) probe, is 62 times more expensive to produce than the Subak reporter.

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