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[Effects of RNA interference on the porcine NHEJ pathway repair factors on HR efficiency].
Yi Chuan ; 40(9): 749-757, 2018 Sep 20.
Artigo em Chinês | MEDLINE | ID: mdl-30369478
Non-homologous end-joining (NHEJ) is the predominant DNA double-strand break (DSB) repair pathway in mammalian cells. It inhibits the efficiency of homologous recombination (HR) by competing for DSB targets. To improve the efficiency of HR in porcine fetal fibroblasts (PFFs), several RNA interference (RNAi) systems were designed to knockdown NHEJ key molecules, such as polynucleotide kinase/phosphatase (PNKP), DNA ligase IV (LIG4) and NHEJ1. The results show that siRNA significantly knocked down LIG4, PNKP and NHEJ1 expression. Suppression of PNKP dramatically increased the efficiency of single-strand annealing (SSA), double-strand DNA (dsDNA) and single-strand DNA (ssODN) mediated homology-directed repair (HDR) by 55.7%, 37.4% and 73.1% after transfected with the SSA-GFP reporter, HDR-GFP system or ssODN-GFP system, respectively; whereas knockdown of LIG4 and NHEJ1 repair factors significantly increased dsDNA or ssODN-mediated HDR efficiency by 37.5% and 76.9%, respectively.
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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: Suínos / Interferência de RNA / Reparo do DNA por Junção de Extremidades / Recombinação Homóloga Limite: Animais Idioma: Chinês Revista: Yi Chuan Assunto da revista: Genética Ano de publicação: 2018 Tipo de documento: Artigo País de afiliação: China