摘要

Frameshift mutations, responsible for >20% of Mendelian inherited diseases, pose substantial therapeutic challenges. Here, we developed Template-Independent Genome Editing for Restoration (TIGER), a platform for efficient and precise correction of frameshift mutations across various models. By identifying reproducible nucleotide-level factors that influence therapeutic efficacy across cells and tissues, we developed a scoring system for guide RNA (gRNA)-Cas9 outcomes. Approximately 75% of deletion and 50% of insertion mutations produced ≥30% in-frame products, sufficient for phenotypic restoration, with 38% and 65% achieving wild-type correction, respectively. To expand TIGER’s applicability across species and genome-wide, we retrained the inDelphi algorithm to predict therapeutic gRNAs for single-nucleotide frameshifts. Validation in a mouse model of deafness, using SpCas9 and optimal gRNA delivered via dual adeno-associated virus, restored hearing thresholds to wild-type levels, with ~90% of in-frame edits being wild-type. TIGER provides a robust and broadly applicable strategy for in vivo correction of inherited frameshift diseases.

个人简介
熊巍博士，北京脑科学与类脑研究所研究员、博士生导师。熊巍博士团队致力于研究听觉系统的分子和神经机制，特别是听觉毛细胞机械转导的分子和细胞机制，言语交流的神经机制，以及耳聋等听觉交流疾病的生物学治疗。近5年来相关工作以通讯作者在PNAS、Neuron、Cell Reports和eLife等杂志上发表。