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Efficient human genome editing using SaCas9 ribonucleoprotein complexes.

08:00 EDT 30th March 2019 | BioPortfolio

Summary of "Efficient human genome editing using SaCas9 ribonucleoprotein complexes."

Genome editing using RNA-guided nucleases in their ribonucleoprotein (RNP) form represents a promising strategy for gene modification and therapy because they are free of exogenous DNA integration and have reduced toxicity in vivo and ex vivo. However, genome editing by Cas9 nuclease from Staphylococcus aureus (SaCas9), has not been reported in its RNP form, which recognizes a longer protospacer adjacent motif (PAM), 5'-NNGRRT-3', compared with SpCas9(Streptococcus pyogenes Cas9) of 5'-NGG-3' PAM. Here, we report SaCas9-RNP mediated genome editing in human cells. The SaCas9-RNP displayed efficient genome editing activities of enhanced green fluorescent protein (EGFP) coding gene as well as three endogenous genes (OPA1, RS1 and VEGFA). We further successfully implemented SaCas9-RNP to correct a pathogenic RS1 mutation for X-linked Juvenile Retinoschisis. We also showed that off-target effects triggered by SaCas9-RNP were undetectable by targeted deep sequencing. Collectively, this study demonstrates the potential of SaCas9-RNP mediated genome editing in human cells, which could facilitate genome editing based therapy. This article is protected by copyright. All rights reserved.

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This article was published in the following journal.

Name: Biotechnology journal
ISSN: 1860-7314
Pages: e1800689

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