TALENs编辑绵羊成纤维细胞FGF 5基因

doi:10.11843/j.issn.0366-6964.2015.05.004

Abstract

Abstract:

Transcriptional activator-like effector （TALE） technologies were established over the last decade as useful tools for genomic editing and transcriptional modulation.Moreover，TALE nucleases (TALENs) technology can be used to bring about targeted gene deletion，insertion and mutagenesis in a siries of species genome.TALENs were designed to target the ATG initiation codon site of ovine FGF 5 gene.According to the results of comparing normal culture and genome editing ovine fibroblasts，the activities of TALEN pairs were tested and screened by Surveyor nuclease after electric transfection TALEN plasmid pairs into ovine fibroblasts.After ovine fibroblast clones were cultured by limiting dilution，the screening of mutant cells was identified by PAGE electrophoresis after PCR amplification.The deletion mutations were introduced into ovine FGF 5 gene of fibroblasts and identified by DNA sequencing.Mutated cells missing the ATG initiation codon of ovine FGF 5 were obtained by valid TALENs.The acquisition of a pair of valid TALENs provides the basis for targeted disruption ovine FGF 5 gene.A point mutation was identified at 104 base site of the ATG initiation codon upstream of ovine FGF 5 gene by Surveyor mutation detection and sequence analysis.

CLC Number:

S826.2

PI Wen-hui，ZHOU Ping，WANG Li-min，TANG Hong，GUO Yan-hua，ZHANG Yi-yuan，LIU Shou-ren，WANG Xin-hua. Editing Fibroblast Growth Factor 5 Gene in Ovine Fibroblasts Using TALENs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(5): 704-710.

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Editing Fibroblast Growth Factor 5 Gene in Ovine Fibroblasts Using TALENs

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