Products/Services Used | Details | Operation |
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Codon Optimization> | The mSpCas9-MLS construct was synthesized by GenScript (NJ, USA) using their OptimumGene™ - Codon Optimization algorithm. | Get A Quote |
Gene editing of the mitochondrial genome using CRISPR-Cas9 system is highly challenging mainly due to sub-efficient delivery of guide RNA and Cas9 enzyme complexes into mitochondria. In this study, we were able to perform gene editing in the mitochondrial DNA by appending NADH-ubiquinone oxidoreductase chain 4 (ND4) targeting guide RNA to a RNA transport derived stem loop element (RP-loop) and expressing the Cas9 enzyme with preceding mitochondrial localization sequence. Our results showed mitochondrial co-localization of RP-loop gRNA and a marked reduction of ND4 expression in the cells carrying a A11204G variant in their ND4 sequence coincidently decreasing the mtDNA levels. This proof-of-concept study sugges... More