Unknown

Dataset Information

0

One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering.


ABSTRACT: 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.

SUBMITTER: Wang H 

PROVIDER: S-EPMC3969854 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature

altmetric image

Publications

One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering.

Wang Haoyi H   Yang Hui H   Shivalila Chikdu S CS   Dawlaty Meelad M MM   Cheng Albert W AW   Zhang Feng F   Jaenisch Rudolf R  

Cell 20130502 4


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 hi  ...[more]

Similar Datasets

| S-EPMC3961003 | biostudies-other
| S-EPMC5518993 | biostudies-literature
| S-EPMC7757693 | biostudies-literature
| S-EPMC5930688 | biostudies-literature
| S-EPMC4973257 | biostudies-literature
| S-EPMC6138433 | biostudies-literature
| S-EPMC8516739 | biostudies-literature
| S-EPMC5713256 | biostudies-literature
| S-EPMC3795411 | biostudies-literature
| S-EPMC6024849 | biostudies-literature