Inhibition of KU70 and KU80 by CRISPR interference, not NgAgo interference, increases the efficiency of homologous recombination in pig fetal fibroblasts
| Autor: | Haoqiang Wang, Huaqiang Yang, Ruan Xiaofang, Jian-xin Mo, Gengyuan Cai, Xianwei Zhang, Rong Quan, Ting Gu, Guo-ling Li, Zhen-fang Wu, Cuili Zhong, Zi-cong Li, Dewu Liu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Ku80 Agriculture (General) homologous recombination Plant Science 01 natural sciences Biochemistry non-homologous end-joining S1-972 chemistry.chemical_compound Food Animals KU80 chemistry.chemical_classification CRISPR interference Gene knockdown Ku70 DNA ligase Ecology Cas9 Chemistry fungi CRISPRi 04 agricultural and veterinary sciences Cell biology 040103 agronomy & agriculture 0401 agriculture forestry and fisheries Animal Science and Zoology NgAgoi KU70 Homologous recombination Agronomy and Crop Science DNA 010606 plant biology & botany Food Science |
| Zdroj: | Journal of Integrative Agriculture, Vol 18, Iss 2, Pp 438-448 (2019) |
| ISSN: | 2095-3119 |
| Popis: | Non-homologous end-joining (NHEJ) is a predominant pathway for the repair of DNA double-strand breaks (DSB). It inhibits the efficiency of homologous recombination (HR) by competing for DSB targets. To improve the efficiency of HR, multiple CRISPR interference (CRISPRi) and Natronobacterium gregoryi Argonaute (NgAgo) interference (NgAgoi) systems have been designed for the knockdown of NHEJ key molecules, KU70, KU80, polynucleotide kinase/phosphatase (PNKP), DNA ligase IV (LIG4), and NHEJ1. Suppression of KU70 and KU80 by CRISPRi dramatically promoted (P0.05) HR efficiency. Interestingly, although the NgAgoi system significantly suppressed (P0.05) HR efficiency in primary fetal fibroblasts. Our result showed that both NgAgo and catalytically inactive Cas9 (dCas9) could interfere with the expression of target genes, but the downstream factors appear to be more active following CRISPR-mediated interference than that of NgAgo. |
| Databáze: | OpenAIRE |
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