CRISPR/Cas9 - Mediated Precise Targeted Integration In Vivo Using a Double Cut Donor with Short Homology Arms

EBioMedicine
Xuan YaoHui Yang

Abstract

Precisely targeted genome editing is highly desired for clinical applications. However, the widely used homology-directed repair (HDR)-based genome editing strategies remain inefficient for certain in vivo applications. We here demonstrate a microhomology-mediated end-joining (MMEJ)-based strategy for precisely targeted gene integration in transfected neurons and hepatocytes in vivo with efficiencies up to 20%, much higher (up to 10 fold) than HDR-based strategy in adult mouse tissues. As a proof of concept of its therapeutic potential, we demonstrate the efficacy of MMEJ-based strategy in correction of Fah mutation and rescue of Fah-/-liver failure mice, offering an efficient approach for precisely targeted gene therapies.

Citations

Aug 23, 2017·Cell Biology and Toxicology·Tetsushi Sakuma, Takashi Yamamoto
Dec 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Alexandre PaixGeraldine Seydoux
Dec 8, 2017·Journal of Human Genetics·Keiichiro Suzuki, Juan Carlos Izpisua Belmonte
Sep 18, 2020·Nucleic Acids Research·Do Yon KimDaesik Kim
Aug 18, 2018·Nature Communications·Shota NakadeTakashi Yamamoto
Aug 26, 2020·Scientific Reports·Riki IshibashiFumiko Toyoshima
Apr 21, 2018·International Journal of Molecular Sciences·Davide De RoccoFiorentina Ascenzioni
Jul 31, 2018·Stem Cells International·Xiangjun HeBo Feng
Jul 12, 2020·Experimental & Molecular Medicine·Hyeon-Ki JangSangsu Bae
Dec 22, 2017·Journal of Cellular Biochemistry·Rajagopal N Aravalli, Clifford J Steer
Oct 14, 2020·Plant Biotechnology Journal·Tien Van VuJae-Yean Kim
May 27, 2020·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Jordan L SmithWen Xue
Nov 17, 2020·Frontiers in Neuroscience·Fábio Duarte, Nicole Déglon
Feb 5, 2021·Journal of Clinical Medicine·Christi T Salisbury-Ruf, Andre Larochelle
Dec 12, 2020·Nucleic Acids Research·Gabriel Martínez-GálvezStephen C Ekker
Aug 28, 2019·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Jun YaoYifan Dai
Feb 20, 2021·Frontiers in Genetics·Iuri Viotti PerisseIrina A Polejaeva
Jun 10, 2018·Clinics and Research in Hepatology and Gastroenterology·Robin LoeschSabine Colnot
Apr 10, 2021·G3 : Genes - Genomes - Genetics·Irina SizovaPeter Hegemann
May 15, 2018·Analytical Chemistry·Kyle J SeamonBrooke Harmon
Sep 13, 2021·Molecular Therapy : the Journal of the American Society of Gene Therapy·Adrian Pickar-OliverCharles A Gersbach

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Methods Mentioned

BETA
gene knock-ins
FACS
PCR
electrophoresis
Infection
Transfection
Genotyping
dissection

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