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BigField GEG Tech's insight:
The authors generated the floxed alleles in rats by CRISPR/Cas9 with an efficiency up to 54%. Then, by crossing Nestin-Cre rat with the Dnmt3b floxed rat and Cck-Cre with Dnmt1 floxed rat, they detected the Cre/loxP-mediated recombination in the F1 generation of rats These results provided a simple and flexible engineering strategy for establishment of knockin rats.
PLOS ONE: an inclusive, peer-reviewed, open-access resource from the PUBLIC LIBRARY OF SCIENCE. Reports of well-performed scientific studies from all disciplines freely available to the whole world.
BigField GEG Tech's insight:
Lentiviral vectors which express shRNA to study cardiac features.
http://geg-tech.com/rna-interference.html
BigField GEG Tech's curator insight,
July 4, 2014 8:14 AM
Lentiviral vectors which express shRNA to study cardiac features.
BigField GEG Tech's insight:
ZFN as a promising tool to manipulate the pig genome. This opens the way of fast generation of KO or KI for pigs.
BigField GEG Tech's insight:
Interesting strategy to produce transgenic mice. The authors estimated an efficiency more than 42%.
BigField GEG Tech's insight:
Original approach of this lab which produces transgenic pigs to design xenografts for human.
researchers_achieved_precise_gene_modification_in_monkeys_monkeys_are_important_for_modeling_diseases_because_of_their_close_similarities_to_humans_but_past_efforts_to_precisely_modify_genes_in_primates_have_failed_in_a_study_published_by_cell_press_january_30th
BigField GEG Tech's insight:
BigField GEG Tech's insight:
An exemple of the power CRISPR system. This type of data makes of CRISPR very attractive tools for transgenesis!
BigField GEG Tech's curator insight,
December 5, 2013 9:02 AM
An exemple of the power CRISPR system. These type of data make of CRISPR very attractive tools for transgenesis!
The "new genetics" promises to change faulty genes of future generations by introducing new, functioning genes using "designer sperm." Research shows that introducing genetic material via a viral vector into mouse sperm leads to the presence and...
BigField GEG Tech's insight:
The lentiviral transduction of spermatozoa, an original and efficient strategy to generate transgenic models!
BigField GEG Tech's curator insight,
December 5, 2013 8:59 AM
The lentiviral transduction of spermatozoa, an original and efficient strategy to generate transgenic models!
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BigField GEG Tech's insight:
The authors provide a detailed protocol for embryo manipulation by piezo-driven injection of nucleic acids into the cytoplasm to create gene-modified mice. Beginning with target design, the generation of gene-modified mice can be achieved in as little as 4 weeks.
SRB's curator insight,
November 1, 2014 7:57 PM
Protocol for Cas9 gene editing in mice from Jaenisch Lab
PMID: 25058643
BigField GEG Tech's insight:
The authors showed that, compared to the TALEN system, the CRISPR/Cas system induced the limb-defect phenotypes with a strikingly higher efficiency. They concluded that although both gene-targeting technologies are useful, the CRISPR/Cas system more effectively elicits single-step biallelic mutations in mice.
BigField GEG Tech's insight:
By microinjecting gRNA, hCas9 mRNA and single-stranded donor oligonucleotides (ssODN) into mouse zygotes, the authors introduced defined genomic modifications in the genomes of mice with a low cost and in a short time.
Humanized mouse models have become increasingly important and widely used in modeling human diseases in biomedical research. Immunodeficient mice such as NOD-Rag1-/-IL2RgammaC-null (NRG) or NOD-SCID-IL2RgammaC-null (NSG) mice are critical for efficient engraftment of human cells or tissues. However, their genetic modification remains challenging due to a lack of embryonic stem cells and difficulty in the collection of timed embryos after superovulation. Here, we report the generation of gene knockout NRG mice by combining in vitro fertilization (IVF) and CRISPR/Cas9 technology. Sufficient numbers of fertilized embryos were produced through IVF, and a high rate of Fah gene targeting was achieved with microinjection of Cas9 mRNA, gRNA and single strand oligonucleotide DNA (ssDNA) into the embryos. The technology paves the way to construct NRG or NSG mutant mice to facilitate new humanized mouse models. The technology can also be readily adapted to introduce mutations in other species such as swine and non-human primates.
BigField GEG Tech's insight:
PLOS ONE: an inclusive, peer-reviewed, open-access resource from the PUBLIC LIBRARY OF SCIENCE. Reports of well-performed scientific studies from all disciplines freely available to the whole world.
BigField GEG Tech's insight:
Another example of the use of lentiviral vector to transduce sperm for the generation of transgenic pigs. The authors estimated an efficiency more than 10%.
BigField GEG Tech's insight:
The authors have designed cytoplasmic injection assays to generate transgenic pigs thanks to the piggybac transoposase. More than 8.00% of the injected embryos developed into transgenic animals containing monogenic and often single transgenes in their genome.
BigField GEG Tech's insight:
Anticipated results: the efficiency of Sleeping Beauty to generate transgenic rabbits (transgenic founders in relation to all animals born alive) should be 15% or higher. The germline transmission from F0 to F1 to be 100%, with transgene segregation close to Mendelian rates (44%).
PLOS ONE: an inclusive, peer-reviewed, open-access resource from the PUBLIC LIBRARY OF SCIENCE. Reports of well-performed scientific studies from all disciplines freely available to the whole world.
BigField GEG Tech's insight:
Background: Environmental endocrine disruptors (EED) are exogenous chemicals that mimic endogenous hormones, such as estrogens. Previous studies using (Transgenic zebrafish reveal tissue-specific differences in estrogen signaling in...
BigField GEG Tech's insight:
BigField GEG Tech's insight:
The combination between transposon and nuclear transfer technique to generate transgenic pigs. Nice approach!
BigField GEG Tech's insight:
An exemple of the power CRISPR system. These type of data make of CRISPR very attractive tools for transgenesis!
BigField GEG Tech's curator insight,
December 5, 2013 9:12 AM
An exemple of the power CRISPR system. This type of data makes of CRISPR very attractive tools for transgenesis!
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