Establishing targeted carp TLR22 gene disruption via homologous recombination using CRISPR/Cas9
Establishing targeted carp TLR22 gene disruption via homologous recombination using CRISPR/Cas9
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DOI：
10.1016/j.dci.2016.04.009
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发表时间：
2016-08-01
影响因子：
2.9
通讯作者：
Barman, Hirak Kumar
中科院分区：
文献类型：
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作者：
Chakrapani, Vemulawada;Patra, Swagat Kumar;Barman, Hirak Kumar
基因编辑技术的最新进展尚未在养殖鱼类中得到利用。我们在养殖鲤鱼Labeo rohita（俗称罗虎）中使用CRISPR/Cas9系统建立了一种基因靶向技术。我们证明了供体DNA是通过同源重组（HR）在CRISPR/Cas9核酸酶产生的靶向双链缺口位点上整合的。这导致成功破坏rohu toll样受体22 （TLR22）基因，该基因参与先天免疫，仅存在于硬骨鱼和两栖动物中。因此产生的零突变体缺乏TLR22 mRNA的表达。总之，这是第一个证据，证明CRISPR/Cas9系统是一种高效的工具，可以通过硬骨鱼的HR靶向基因破坏，产生大型养殖鱼类模型。(C) 2016 Elsevier Ltd.版权所有。
Recent advances in gene editing techniques have not been exploited in farmed fishes. We established a gene targeting technique, using the CRISPR/Cas9 system in Labeo rohita, a farmed carp (known as rohu). We demonstrated that donor DNA was integrated via homologous recombination (HR) at the site of targeted double-stranded nicks created by CRISPR/Cas9 nuclease. This resulted in the successful disruption of rohu Toll-like receptor 22 (TLR22) gene, involved in innate immunity and exclusively present in teleost fishes and amphibians. The null mutant, thus, generated lacked TLR22 mRNA expression. Altogether, this is the first evidence that the CRISPR/Cas9 system is a highly efficient tool for targeted gene disruption via HR in teleosts for generating model large-bodied farmed fishes. (C) 2016 Elsevier Ltd. All rights reserved.