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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 910-918 |
| Number of pages | 9 |
| Journal | Cell |
| Volume | 153 |
| Issue number | 4 |
| DOIs | |
| State | Published - May 9 2013 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
One-step generation of mice carrying mutations in multiple genes by CRISPR/cas-mediated genome engineering. / Wang, Haoyi; Yang, Hui; Shivalila, Chikdu S.; Dawlaty, Meelad M.; Cheng, Albert W.; Zhang, Feng; Jaenisch, Rudolf.
In: Cell, Vol. 153, No. 4, 09.05.2013, p. 910-918.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - One-step generation of mice carrying mutations in multiple genes by CRISPR/cas-mediated genome engineering
AU - Wang, Haoyi
AU - Yang, Hui
AU - Shivalila, Chikdu S.
AU - Dawlaty, Meelad M.
AU - Cheng, Albert W.
AU - Zhang, Feng
AU - Jaenisch, Rudolf
PY - 2013/5/9
Y1 - 2013/5/9
N2 - 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.
AB - 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.
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U2 - 10.1016/j.cell.2013.04.025
DO - 10.1016/j.cell.2013.04.025
M3 - Article
VL - 153
SP - 910
EP - 918
JO - Cell
JF - Cell
SN - 0092-8674
IS - 4
ER -