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Rescooped by wb from CRISPR-Cas System for Eukaryotic Genome Engineering
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Genome editing in human pluripotent stem cells | StemBook

Genome editing in human pluripotent stem cells | StemBook | genome modifications | Scoop.it

Genome editing is used to make targeted modifications to the genome of eukaryotic cells. There are many potential applications of genome editing in human pluripotent stem cells (hPSCs) including the generation of knockout and reporter cell lines. This protocol describes a system for efficient genome editing in hPSCs using engineered transcription activator-like effector nucleases (TALENs) or clustered regularly interspaced short palindromic repeat (CRISPR) technology.


Via Amir Taheri Ghahfarokhi
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Jun Liu's curator insight, July 4, 2013 11:50 PM

TALEN libray!

Rescooped by wb from CRISPR-Cas System for Eukaryotic Genome Engineering
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One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering

One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering | genome modifications | Scoop.it

Wang et al., 2013, Cell

 

Highlights

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> CRISPR/Cas9-mediated simultaneous targeting of five genes in mES cells

> Generation of Tet1/Tet2 double-mutant mice in one step

> Generation of Tet1/Tet2 double-mutant mice with predefined mutations in one step

 

Summary

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Mice carrying mutations in multiple genes are traditionally generated by sequential recombination in embryonic stem cells and/or time-consuming intercrossing of mice with a single mutation. The CRISPR/Cas system has been adapted as an efficient gene-targeting technology with the potential for multiplexed genome editing. We demonstrate that CRISPR/Cas-mediated gene editing allows the simultaneous disruption of five genes (Tet1, 2, 3, Sry, Uty - 8 alleles) in mouse embryonic stem (ES) cells with high efficiency. Coinjection of Cas9 mRNA and single-guide RNAs (sgRNAs) targeting Tet1 and Tet2 into zygotes generated mice with biallelic mutations in both genes with an efficiency of 80%. Finally, we show that coinjection of Cas9 mRNA/sgRNAs with mutant oligos generated precise point mutations simultaneously in two target genes. Thus, the CRISPR/Cas system allows the one-step generation of animals carrying mutations in multiple genes, an approach that will greatly accelerate the in vivo study of functionally redundant genes and of epistatic gene interactions.


Via Amir Taheri Ghahfarokhi
wb's insight:

amazing.....

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Rescooped by wb from CRISPR-Cas System for Eukaryotic Genome Engineering
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CRISPR Genome Engineering Resources | learn, share, and discuss

CRISPR Genome Engineering Resources | learn, share, and discuss | genome modifications | Scoop.it

Prof. Feng Zhang CRISPR/Cas system



Via Amir Taheri Ghahfarokhi
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High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells

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too high off target

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Rescooped by wb from Multi- gene
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Transgenics: A new breed - Nature.com

Transgenics: A new breed - Nature.com | genome modifications | Scoop.it
Nature.com
Transgenics: A new breed
Nature.com
For example, enzymes called transcription activator-like effector nucleases (TALENs) and zinc-finger nucleases (ZFNs) can cut DNA at specific points chosen by the experimenter.

Via Christian Faltado Cantos
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