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Improvement of TaC9-ABE mediated correction of human SMN2 gene
  • +9
  • Min Chen,
  • Xiaohua Peng,
  • Yue Chi,
  • Jinling Wang,
  • Shuangpeng Li,
  • Yang liu,
  • Chengcheng Tang,
  • Xiaoqing Zhou,
  • Xuan Lu,
  • Yue Gao,
  • Liangxue Lai,
  • Qingjian Zou
Min Chen
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Xiaohua Peng
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Yue Chi
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Jinling Wang
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Shuangpeng Li
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Yang liu
BGI-Hangzhou
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Chengcheng Tang
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Xiaoqing Zhou
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Xuan Lu
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Yue Gao
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Liangxue Lai
South China Institute of Large Animal Models for Biomedicine Wuyi University
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Qingjian Zou
South China Institute of Large Animal Models for Biomedicine Wuyi University

Corresponding Author:[email protected]

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Abstract

Spinal muscular atrophy (SMA) is a devastating neuromuscular disease caused by mutations in the survival motor neuron 1 ( SMN1) gene. Gene editing technology repairs the conversion of the 6th base T to C in exon 7 of the paralogous SMN2 gene, compensating for the SMN protein expression and promoting the survival and function of motor neurons. However, low editing efficiency and unintended off-target effects limit the application of this technology. Here, we optimized a TaC9-adenine base editor (ABE) system by combining Cas9 nickase with the transcription activator-like effector (TALE)-adenosine deaminase fusion protein to effectively and precisely edit SMN2 without detectable Cas9 dependent off-target effects in human cell lines. We also generated human SMA-induced pluripotent stem cells (SMA-iPSCs) through the mutation of the splice acceptor or deletion of the exon 7 of SMN1. TaC9-R10 induced 45% SMN2 T6>C conversion in the SMA-iPSCs. The SMN2 T6>C splice-corrected SMA-iPSCs were directionally differentiated into motor neurons, exhibiting SMN protein recovery and anti-apoptosis ability. Therefore, the TaC9-ABE system with dual guides from the combination of Cas9 with TALE could be a potential therapeutic strategy for SMA with high efficacy and safety.