Homology-directed repair in rodent zygotes using Cas9 and TALEN engineered proteins

被引：63

作者：

Menoret, Severine ^{[1
,2
,3
]}

De Cian, Anne ^{[4
]}

Tesson, Laurent ^{[1
,2
,3
]}

Remy, Severine ^{[1
,2
,3
]}

Usal, Claire ^{[1
,2
,3
]}

Boule, Jean-Baptiste ^{[4
]}

Boix, Charlotte ^{[4
]}

Fontaniere, Sandra ^{[5
]}

Creneguy, Alison ^{[1
,2
]}

Nguyen, Tuan H. ^{[1
,2
]}

Brusselle, Lucas ^{[1
,2
]}

Thinard, Reynald ^{[1
,2
,3
]}

Gauguier, Dominique ^{[6
,7
]}

Concordet, Jean-Paul ^{[4
]}

Cherifi, Yacine ^{[5
]}

Fraichard, Alexandre ^{[5
]}

Giovannangeli, Carine ^{[4
]}

Anegon, Ignacio ^{[1
,2
,3
]}

机构：

[1] INSERM, UMR 1064 ITUN, F-44093 Nantes, France

[2] CHU Nantes, F-44093 Nantes, France

[3] CNRS, UMS3556, SFR Francois Bonamy, Platform Rat Transgenesis Immunophen, F-44093 Nantes, France

[4] Sorbonne Univ, Museum Natl Hist Nat, Struct & Instabil Genomes, INSERM,U1154,CNRS UMR7196, F-75005 Paris, France

[5] genOway, F-69007 Lyon, France

[6] Univ Paris 05, Univ Paris 06, Sorbonne Paris Cite,Cordeliers Res Ctr, Sorbonne Univ,INSERM,UMR S 1138, F-75006 Paris, France

[7] Univ Paris 06, Inst Cardiometab & Nutr, F-75013 Paris, France

来源：

SCIENTIFIC REPORTS | 2015年 / 5卷

关键词：

GENE KNOCKOUT; ZINC-FINGER; CONDITIONAL ALLELES; DIRECT-INJECTION; RAT; GENERATION; MICE; MOUSE; MUTATIONS; DELIVERY;

D O I：

10.1038/srep14410

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The generation of genetically-modified organisms has been revolutionized by the development of new genome editing technologies based on the use of gene-specific nucleases, such as meganucleases, ZFNs, TALENs and CRISPRs-Cas9 systems. The most rapid and cost-effective way to generate genetically-modified animals is by microinjection of the nucleic acids encoding gene-specific nucleases into zygotes. However, the efficiency of the procedure can still be improved. In this work we aim to increase the efficiency of CRISPRs-Cas9 and TALENs homology-directed repair by using TALENs and Cas9 proteins, instead of mRNA, microinjected into rat and mouse zygotes along with long or short donor DNAs. We observed that Cas9 protein was more efficient at homology-directed repair than mRNA, while TALEN protein was less efficient than mRNA at inducing homology-directed repair. Our results indicate that the use of Cas9 protein could represent a simple and practical methodological alternative to Cas9 mRNA in the generation of genetically-modified rats and mice as well as probably some other mammals.

引用

页数：15

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