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Efficient Intracellular Delivery of CRISPR-Cas Ribonucleoproteins through Receptor Mediated Endocytosis


We recently reported a new delivery system harnessing surface receptors for targeted uptake of CRISPR-Cas9 ribonucleoprotein into mammalian cells (Rouet et al., JACS 2018). For this purpose, Cas9 protein was labeled with the small molecule ligand ASGRL, specific for the asialoglycoprotein receptor, enabling endosomal uptake of the ribonucleoprotein into human cells expressing the receptor. However, detailed mechanistic insights had remained unknown and editing efficiency low. Here we investigate the mechanism of endosomal escape as mediated by the ppTG21 endosomolytic peptide and outline the development of novel Cas9 or Cas12a ribonucleoprotein complexes with increased editing efficiency.

Type Journal
ISBN 1554-8937 (Electronic) 1554-8929 (Linking)
Authors Rouet, R.; Christ, D.
Responsible Garvan Author (missing name)
Publisher Name ACS Chemical Biology
Published Date 2019-03-15
Published Volume 14
Published Issue 3
Published Pages 554-561
Status Published in-print
DOI 10.1021/acschembio.9b00116
URL link to publisher's version