Microenvironment-Responsive Delivery of the Cas9 RNA-Guided Endonuclease for Efficient Genome Editing

Bioconjugate Chemistry
Jun YinXiangdong Gao

Abstract

Successful and efficient delivery of Cas9 protein and gRNA into cells is critical for genome editing and its therapeutic application. In this study, we developed an improved supercharged polypeptide (SCP) mediated delivery system based on dithiocyclopeptide linker to realize the effective genome editing in tumor cells. The fusion protein Cas9-linker-SCP (Cas9-LS) forms positively charged complexes with gRNA in vitro to provide possibilities for gRNA delivery into cells. Under the microenvironment of tumor cells, the dithiocyclopeptide linker, containing matrix metalloproteinase 2 (MMP-2) sensitive sequence and an intramolecular disulfide bond, can be completely disconnected to promote the release of Cas9 protein with the nuclear localization sequence (NLS) in the cytoplasm and transfer to the cell nucleus for highly efficient genome editing, resulting in an obvious increase of indel efficiency in comparison to fusion protein without dithiocyclopeptide linker (Cas9-SCP). Furthermore, Cas9-LS shows no significant cytotoxicity and minimal hemolytic activity. We envision that the microenvironment-responsive Cas9 protein delivery system can facilitate more efficient genome editing in tumor cells.

References

May 24, 2001·Immunity·H Wagner
Jan 9, 2010·Science·Philippe Horvath, Rodolphe Barrangou
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jul 4, 2012·Nature Methods·Thomas GajCarlos F Barbas
Jan 31, 2013·Nature Biotechnology·Wenyan JiangLuciano A Marraffini
Jan 31, 2013·Nature Biotechnology·Seung Woo ChoJin-Soo Kim
Mar 13, 2013·Biomaterials·Diego PesceAndreas Herrmann
Jun 25, 2013·Nature Biotechnology·Yanfang FuJeffry D Sander
Apr 1, 2014·Nature Biotechnology·Hao YinDaniel G Anderson
May 25, 2015·Journal of Biotechnology·Xiquan LiangJonathan D Chesnut
Oct 31, 2015·Journal of Controlled Release : Official Journal of the Controlled Release Society·Huining HeVictor C Yang
Mar 2, 2016·Proceedings of the National Academy of Sciences of the United States of America·Ming WangQiaobing Xu
Oct 14, 2016·Nature·Martin P StewartKlavs F Jensen
Aug 2, 2017·Nature Plants·Kangquan YinJin-Long Qiu
Sep 16, 2017·Journal of Controlled Release : Official Journal of the Controlled Release Society·Chang LiuKun Cheng
Sep 28, 2017·Nature·Norah M E FogartyKathy K Niakan
Jan 7, 2018·Trends in Biotechnology·Zachary GlassQiaobing Xu
May 26, 2018·ACS Applied Materials & Interfaces·Guojun ChenShaoqin Gong
Nov 7, 2018·Journal of the American Chemical Society·Jun YinXiangdong Gao

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Citations

Aug 9, 2021·Journal of Controlled Release : Official Journal of the Controlled Release Society·Bixi SunXiaoshu Gao
Jan 15, 2020·Journal of the American Chemical Society·Mram Z AlyamiNiveen M Khashab
Dec 3, 2021·Biomacromolecules·Justin M Horn, Allie C Obermeyer
Jan 19, 2022·ACS Applied Bio Materials·Akbar HasanzadehMichael R Hamblin
Feb 2, 2022·Biomaterials Science·Yi LinUlrich Lächelt

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