利用TALENs技术建立基因定点修饰的猪Oct-4-EGFP细胞系

Abstract

Abstract: This study aims to establish a reporting system for stem cells tracer, EGFP cDNA together with LoxP-flanked PGK-neo cassettes were inseet into the Oct-4 gene of Wuzhishan Miniature pig at the stop codon via TALENs mediated homologous recombination, expression of EGFP gene was under control of Oct-4 5'-regulatory region. TALENs directed against sequences flanking the stop codon of Oct-4 were cotrasfected with targeting vector, 514 drug-resistant cell clones were screened, and 36 correctly targeted clones were confirmed by PCR from them, the targeting efficiency were 5.6% and 13.0%. Oct-4-EGFP transgenic cell line was establinshed in this study, and it was also proved that TALENs could significantly improve the efficiency of homologous recombination.

Key words: Oct-4-EGFP; TALENs; homologous recombination; pig; stem cell; pluripotency

CLC Number:

Q813

LIU Chang, FENG Chong, SONG Zhi-qiang, LI Xi-rui, WANG Ning, CHU Ming-xing, PAN Deng-ke. Estblishment of Porcine Oct-4-EGFP Site-directed Transgene Cell Lines using Engineered TALENs[J]. , 2013, 40(11): 1-6.

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Estblishment of Porcine Oct-4-EGFP Site-directed Transgene Cell Lines using Engineered TALENs

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