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.