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

Autor: Chakrapani V; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India., Patra SK; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India., Panda RP; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India., Rasal KD; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India., Jayasankar P; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India., Barman HK; Fish Genetics and Biotechnology Division, ICAR - Central Institute of Freshwater Aquaculture, Bhubaneswar 751 002, Odisha, India. Electronic address: Hirak.Barman@icar.gov.in.
Jazyk: angličtina
Zdroj: Developmental and comparative immunology [Dev Comp Immunol] 2016 Aug; Vol. 61, pp. 242-7. Date of Electronic Publication: 2016 Apr 11.
DOI: 10.1016/j.dci.2016.04.009
Abstrakt: 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.
(Copyright © 2016 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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