Establishing targeted carp TLR22 gene disruption via homologous recombination using CRISPR/Cas9

被引：60

作者：

Chakrapani, Vemulawada ^{[1
]}

Patra, Swagat Kumar ^{[1
]}

Panda, Rudra Prasanna ^{[1
]}

Rasal, Kiran Dashrath ^{[1
]}

Jayasankar, Pallipuram ^{[1
]}

Barman, Hirak Kumar ^{[1
]}

机构：

[1] ICAR Cent Inst Freshwater Aquaculture, Fish Genet & Biotechnol Div, Bhubaneswar 751002, Odisha, India

来源：

DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY | 2016年 / 61卷

关键词：

Carp TLR22; Gene editing; CRISPR/Cas9; Microinjection; Homologous recombination; TOLL-LIKE RECEPTORS; EXPRESSION ANALYSIS; MOLECULAR-CLONING; LABEO-ROHITA; COMPARATIVE RESPONSES; SPERMATOGONIAL CELLS; LIGANDS STIMULATION; 1ST EVIDENCE; ROHU CARP; EFFICIENT;

D O I：

10.1016/j.dci.2016.04.009

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

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.

引用

页码：242 / 247

页数：6

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