三种小分子化合物对猪胎儿成纤维细胞DNA精确修复效率的影响

doi:10.11843/j.issn.0366-6964.2018.12.007

Abstract

Abstract:

The aim of this study was to investigate the effects of 3 small molecule compounds (RS-1, Chir99021 and PD173074) on the DNA precise repair efficiency in porcine fetal fibroblasts. In this study,the activator RS-1 for Rad51, the inhibitor Chir99021 for glycogen synthase kinase 3 (GSK-3) and the inhibitor PD173074 for fibroblast growth factor receptor (FGFR1) were used to culture porcine fetal fibroblasts, and then the percentage of green fluorescent cells was detected by flow cytometry to verify the effect of 3 small molecule compounds on DNA repair efficiency. The results showed that:1) After treatment with different doses of Chir99021 and PD173074, the expression levels of PFFs DNA damage repair factors LIG4, NHEJ1, XRCC5, XRCC6 and Rad51 were significantly down-regulated (P<0.05), and after treatment with different doses of of PD173074, the expression level of PNKP factor was significantly up-regulated (P<0.05).2) The low dose of Chir99021 significantly increased the efficiency of homologous recombination (P<0.05) mediated by homologous recombination (HR) repair(P<0.05). High dose of Chir99021 could significantly improve the efficiency of homologous recombination mediated by single strand annealing (SSA) repair(P<0.05).The low doses of RS-1 and PD173074 significantly increased HR efficiency (P<0.05), but RS-1 significantly down-regulated SSA repair efficiency at high concentration (P<0.05), while PD173074 had no significant effect on SSA efficiency (P>0.05). Furthermore, the single-stranded oligonucleotide-mediated DNA repair (ssODN) was not affected (P>0.05) by small molecule compounds. This study indicates that the application of small molecule compounds with appropriate concentrations will facilitate the generation of gene knock-in cell lines, which laying the foundation for obtaining genetic modification animal models.

CLC Number:

S828.2

LI Guo-ling, WANG Hao-qiang, RUAN Xiao-fang, MO Jian-xin, QUAN Rong, ZHONG Cui-li, LI Zi-cong, WU Zhen-fang, LIU De-wu, ZHANG Xian-wei. Effect of Three Small Molecule Compounds on Precise DNA Repair Efficiency in Porcine Fetal Fibroblasts[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2584-2592.

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Effect of Three Small Molecule Compounds on Precise DNA Repair Efficiency in Porcine Fetal Fibroblasts

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