Genetic Modulation of RNA Splicing with a CRISPR-Guided Cytidine Deaminase
Yunqing Ma, Juanjuan Yuan, Xing Chang
STAR Protocols · 2020
doi:10.1016/j.xpro.2019.100005 (opens in a new tab)
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Abstract
Summary: This protocol uses lipofectamine to deliver base editors (i.e., dCas9 and AIDx fusion protein) and sgRNA expression vectors into Duchenne Muscular Dystrophy (DMD) patient-derived human induced pluripotent stem cells (hiPSCs). This protocol details mutation of the 5′ splice site of DMD exon50 with TAM (targeted AID-induced mutagenesis) followed by amplicon-based NGS library preparation for high-throughput sequencing analysis. This protocol can be generalized for base editing in other hIPSCs and for correcting aberrant splicing associated with other genetic diseases.For complete information on the generation and use of this protocol, please refer to Yuan et al. (2018).
Abstract licence: CC BY-NC
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