A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice

Nature Biotechnology
Yang YangJames M Wilson

Abstract

Many genetic liver diseases in newborns cause repeated, often lethal, metabolic crises. Gene therapy using nonintegrating viruses such as adeno-associated virus (AAV) is not optimal in this setting because the nonintegrating genome is lost as developing hepatocytes proliferate. We reasoned that newborn liver may be an ideal setting for AAV-mediated gene correction using CRISPR-Cas9. Here we intravenously infuse two AAVs, one expressing Cas9 and the other expressing a guide RNA and the donor DNA, into newborn mice with a partial deficiency in the urea cycle disorder enzyme, ornithine transcarbamylase (OTC). This resulted in reversion of the mutation in 10% (6.7-20.1%) of hepatocytes and increased survival in mice challenged with a high-protein diet, which exacerbates disease. Gene correction in adult OTC-deficient mice was lower and accompanied by larger deletions that ablated residual expression from the endogenous OTC gene, leading to diminished protein tolerance and lethal hyperammonemia on a chow diet.

References

Jun 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·P E Hodges, L E Rosenberg
Aug 1, 1996·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·M A DingemanseA F Moorman
Aug 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·Guang-Ping GaoJames M Wilson
Nov 2, 2005·Nature Reviews. Microbiology·Ana Vasileva, Rolf Jessberger
Apr 17, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sharon C CunninghamIan E Alexander
Apr 10, 2009·Current Gene Therapy·Paul A LoDucaRoland W Herzog
May 11, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Yu Fu WangHua-Ping Liang
Aug 4, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Lili WangJames M Wilson
Nov 22, 2011·Human Gene Therapy·Lili WangJames M Wilson
Aug 21, 2012·The Journal of Pediatrics·Nicholas Ah MewUNKNOWN Urea Cycle Disorders Consortium of the Rare Diseases Clinical Research Network
Oct 3, 2013·Blood·Xavier M AnguelaKatherine A High
Oct 26, 2013·Nature Protocols·F Ann RanFeng Zhang
Apr 1, 2014·Nature Biotechnology·Hao YinDaniel G Anderson
Aug 20, 2014·Molecular Genetics and Metabolism·Mark L BatshawUNKNOWN Members of the Urea Cycle Disorders Consortium
Nov 29, 2014·Science·Jennifer A Doudna, Emmanuelle Charpentier
Dec 17, 2014·Nature Biotechnology·Shengdar Q TsaiJ Keith Joung
Feb 11, 2015·Biotechnology Journal·Florian Schmidt, Dirk Grimm
Apr 2, 2015·Nature·F Ann RanFeng Zhang
May 20, 2015·Nature Biotechnology·Azita J MahinyMichael S D Kormann
May 30, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Christian HindererJames M Wilson
Jun 23, 2015·Nature·Benjamin P KleinstiverJ Keith Joung
Jun 1, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sharon C CunninghamIan E Alexander

❮ Previous
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Citations

Apr 1, 2016·Gene Therapy·H-Y XueY Xu
Jun 9, 2016·Journal of Molecular Endocrinology·Suzy Markossian, Frédéric Flamant
Jul 8, 2016·Human Genetics·Channabasavaiah B GurumurthyXue Zhong Liu
Aug 9, 2016·Briefings in Functional Genomics·Tomoko Kato, Shuji Takada
Aug 10, 2016·Nature Biotechnology·Cormac Sheridan
Aug 27, 2016·Briefings in Functional Genomics·Taeyoung Koo, Jin-Soo Kim
May 25, 2016·Annual Review of Genomics and Human Genetics·Xin XiongLei S Qi
Sep 7, 2016·Nature Methods·Wei Leong ChewGeorge M Church
Nov 5, 2016·Nature Reviews. Cardiology·Alanna Strong, Kiran Musunuru
Jan 11, 2017·Hämostaseologie·Simone A HaasToni Cathomen
Jan 12, 2017·Human Gene Therapy·Paul N Valdmanis, Mark A Kay
Apr 8, 2017·Nature Medicine·Tatjana I CornuToni Cathomen
Apr 11, 2017·Acta Pharmacologica Sinica·Gayong ShimYu-Kyoung Oh
Apr 4, 2017·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chaoran YinWenhui Hu
May 21, 2017·Science China. Life Sciences·Zhi-Yao HeYu-Quan Wei
May 31, 2017·ACS Synthetic Biology·Michael PinedaSamira Kiani
Feb 26, 2016·Neuropharmacology·Sourav R ChoudhuryPaola Grandi
May 26, 2017·Journal of Cellular Biochemistry·Xing-Da JuZhong Sheng Sun
Feb 28, 2017·BMC Medicine·Jessica L SchnellerCharles P Venditti
Feb 1, 2017·Current Opinion in Ophthalmology·Thiago CabralStephen H Tsang
Dec 29, 2017·Virus Genes·Feifei WangQin Chen
Mar 9, 2018·The Journal of Clinical Investigation·Amber W WangKlaus H Kaestner
Mar 7, 2018·The Journal of Biological Chemistry·Yanjiao ShaoDali Li
Apr 29, 2018·Biotechnology Letters·Hasan Mollanoori, Shahram Teimourian
May 26, 2017·Journal of Cellular Biochemistry·Vijai SinghChristian Kuete Fofié
Apr 8, 2016·Blood·Matthew C Canver, Stuart H Orkin
Feb 16, 2018·Human Gene Therapy·Yang YangYuquan Wei
Jan 10, 2018·Biotechnology Journal·Carolin Schmelas, Dirk Grimm
Sep 1, 2018·Expert Opinion on Drug Delivery·D C LutherV M Rotello
Jul 29, 2017·Arteriosclerosis, Thrombosis, and Vascular Biology·Alexandra C ChadwickKiran Musunuru
Jul 22, 2018·Arteriosclerosis, Thrombosis, and Vascular Biology·Kelsey E JarrettWilliam R Lagor
Aug 26, 2017·Arteriosclerosis, Thrombosis, and Vascular Biology·Alexandra C Chadwick, Kiran Musunuru
Mar 4, 2017·Circulation Research·Timon SeegerJoseph C Wu
Nov 6, 2018·Chembiochem : a European Journal of Chemical Biology·Le WangXingyu Jiang

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Methods Mentioned

BETA
transfection
PCR
targeted modifications
gene replacement therapy
Assay
Protein assay
Illumina sequencing

Software Mentioned

GraphPad Prism
custom
ImageJ Analyze Particles
ImageLab
custom scripts

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