Ligase IV inhibitor SCR7 enhances gene editing directed by CRISPR-Cas9 and ssODN in human cancer cells

Cell & Bioscience
Zheng HuYuan-Shan Zhu

Abstract

Precise genome editing is essential for both basic and translational research. The recently developed CRISPR/Cas9 system can specifically cleave a designated site of target gene to create a DNA double-strand break, which triggers cellular DNA repair mechanism of either inaccurate non-homologous end joining, or site-specific homologous recombination. Unfortunately, homology-directed repair (HDR) is challenging due to its very low efficiency. Herein, we focused on improving the efficiency of HDR using a combination of CRISPR/Cas9, eGFP, DNA ligase IV inhibitor SCR7, and single-stranded oligodeoxynucleotides (ssODN) in human cancer cells. When Cas9, gRNA and eGFP were assembled into a co-expression vector, the disruption rate more than doubled following GFP-positive cell sorting in transfected cells compared to those unsorted cells. Using ssODNs as templates, SCR7 treatment increased targeted insertion efficiency threefold in transfected cells compared to those without SCR7 treatment. Moreover, this combinatorial approach greatly improved the efficiency of HDR and targeted gene mutation correction at both the GFP-silent mutation and the β-catenin Ser45 deletion mutation cells. The data of this study suggests that a combination of ...Continue Reading

References

Jun 21, 1994·Proceedings of the National Academy of Sciences of the United States of America·P RouetM Jasin
Sep 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·M IlyasW F Bodmer
Jan 30, 2004·Cancer Cell·Hans Clevers
Dec 25, 2007·Cell Research·Meena ShrivastavJac A Nickoloff
Jan 2, 2008·Cell Research·Xuan Li, Wolf-Dietrich Heyer
Dec 10, 2008·Proceedings of the National Academy of Sciences of the United States of America·Kelly J BeumerDana Carroll
Aug 4, 2010·Methods in Molecular Biology·Dmitry Y GuschinEdward J Rebar
Aug 19, 2010·Nature Reviews. Genetics·Fyodor D UrnovPhilip D Gregory
Jul 19, 2011·Nature Methods·Fuqiang ChenGregory D Davis
Jun 12, 2012·Cell·Hans Clevers, Roel Nusse
Jun 20, 2012·Proceedings of the National Academy of Sciences of the United States of America·Hiroshi OchiaiTakashi Yamamoto
Jan 5, 2013·Science·Le CongFeng Zhang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Jan 31, 2013·Nature Biotechnology·Woong Y HwangJ Keith Joung
Jan 31, 2013·Nature Biotechnology·Seung Woo ChoJin-Soo Kim
Mar 27, 2013·Cell Research·Nannan ChangJianzhong Jeff Xi
Apr 3, 2013·Cell Research·Bin ShenXingxu Huang
Oct 25, 2013·PloS One·Daniel Gomez-CabelloPablo Huertas
Dec 10, 2013·Cell Stem Cell·Yuxuan WuJinsong Li
Dec 18, 2013·Science·Tim WangEric S Lander
Dec 18, 2013·Science·Ophir ShalemFeng Zhang
Jan 10, 2014·Development·Xiaogang GuoYonglong Chen
May 6, 2015·Circulation Research·Parwiz AbrahimiJordan S Pober
Aug 14, 2015·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Zhaoying ShiYonglong Chen
Aug 21, 2015·The FEBS Journal·Supriya V Vartak, Sathees C Raghavan
Jan 2, 2016·Science·Mohammadsharif TabebordbarAmy J Wagers
Jan 13, 2016·Cellular and Molecular Life Sciences : CMLS·Yan ZhouYonglun Luo
Mar 11, 2016·Scientific Reports·Jong Seong HaDae-Ro Ahn
Sep 25, 2016·Methods in Molecular Biology·Guillaume Andrey, Malte Spielmann

❮ Previous
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Citations

Oct 26, 2018·Journal of Dental Research·N YuW V Giannobile
Apr 1, 2020·Molecular Reproduction and Development·I Lamas-ToranzoP Bermejo-Álvarez
Jun 6, 2020·Archives of Pharmacal Research·In-Sook JeonJoong-Kook Choi
Sep 10, 2020·International Journal of Molecular Sciences·Han YangNingshao Xia
Dec 4, 2019·Nature Cell Biology·Charles D YehJacob E Corn
Mar 10, 2019·Nature Communications·Grégoire CullotAurélie Bedel
Dec 13, 2019·BioMed Research International·Junjiao WuChun-Li Zhang
Feb 10, 2020·Molecular Biology Reports·A A AnuchinaS A Smirnikhina
Jul 16, 2020·Cells·Antonio Carusillo, Claudio Mussolino
Dec 12, 2020·Computational and Structural Biotechnology Journal·Ainsley Mike AntaoSuresh Ramakrishna
Feb 7, 2021·Biochemical and Biophysical Research Communications·Ben YangTao Li
Sep 28, 2020·Journal of Biotechnology·Mujtaba Aamir BhatArif Tasleem Jan
Apr 27, 2021·Trends in Genetics : TIG·Chaoyou Xue, Eric C Greene
Mar 18, 2020·Genes & Diseases·Zhi-Yong ZhangFang Zhang
Aug 9, 2021·Nucleic Acids Research·Mengzhu LiuJiazhi Hu
Aug 28, 2021·International Journal of Molecular Sciences·Christopher E DenesDaniel Hesselson
Aug 31, 2021·Bio-protocol·Masahito Yamagata, Joshua R Sanes
Aug 18, 2020·Biochemical Pharmacology·Ujjayinee Ray, Sathees C Raghavan

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Methods Mentioned

BETA
genetic modifications
transfection
PCR
fluorescence
electrophoresis
FACS
gene modification
flow cytometry
targeted gene modification

Software Mentioned

SPSS
Image J

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