期刊
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
卷 61, 期 -, 页码 242-247出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.dci.2016.04.009
关键词
Carp TLR22; Gene editing; CRISPR/Cas9; Microinjection; Homologous recombination
资金
- National Agricultural Science Fund (NASF), Indian Council of Agricultural Research, Union Ministry of Agriculture, Government of India [NASF/BS-3017]
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.
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