Improvement of TaC9-ABE mediated correction of human SMN2 gene.
Biotechnol Bioeng
; 121(10): 3059-3067, 2024 Oct.
Article
em En
| MEDLINE
| ID: mdl-38923503
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 antiapoptosis 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.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Proteína 2 de Sobrevivência do Neurônio Motor
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Células-Tronco Pluripotentes Induzidas
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Edição de Genes
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article