Chinese Journal of Comparative Medicine 2014;(3):55-60

doi:10.3969/j.issn.1671.7856.2014.003.012

Generating insulin receptor substrate 1 (Irs1) knockout rat using CRISPR/Cas9

Yuanwu MA ; Jing MA ; Yingdong LU ; Wei CHEN ; Xu ZHANG ; Lei YU ; Lianfeng ZHANG

Keywords

Irs1; Gene knockout; Rat; CRISPR/Cas9

Country

China

Language

Chinese

Abstract

Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .