CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes
Puping Liang,Yanwen Xu,Xiya Zhang,Chenhui Ding,Rui Huang,Zhen Zhang,Jie Lv,Xiaowei Xie,Yuxi Chen,Yujing Li,Sun Ying,Yaofu Bai,Zhou Songyang,Wenbin Ma,Canquan Zhou,Junjiu Huang +15 more
TLDR
It is found that CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB), however, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic.Abstract:
Genome editing tools such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) have been widely used to modify genes in model systems including animal zygotes and human cells, and hold tremendous promise for both basic research and clinical applications. To date, a serious knowledge gap remains in our understanding of DNA repair mechanisms in human early embryos, and in the efficiency and potential off-target effects of using technologies such as CRISPR/Cas9 in human pre-implantation embryos. In this report, we used tripronuclear (3PN) zygotes to further investigate CRISPR/Cas9-mediated gene editing in human cells. We found that CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB). However, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Off-target cleavage was also apparent in these 3PN zygotes as revealed by the T7E1 assay and whole-exome sequencing. Furthermore, the endogenous delta-globin gene (HBD), which is homologous to HBB, competed with exogenous donor oligos to act as the repair template, leading to untoward mutations. Our data also indicated that repair of the HBB locus in these embryos occurred preferentially through the non-crossover HDR pathway. Taken together, our work highlights the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing.read more
Citations
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Journal ArticleDOI
CRISPR/Cas9 in Genome Editing and Beyond
TL;DR: The Cas9 protein, derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is emerging as a powerful tool for engineering the genome in diverse organisms.
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Gene therapy comes of age.
Cynthia E. Dunbar,Katherine A. High,J. Keith Joung,Donald B. Kohn,Keiya Ozawa,Michel Sadelain +5 more
TL;DR: The pioneering work that led the gene therapy field to its current state is reviewed, gene-editing technologies that are expected to play a major role in the field's future are described, and practical challenges in getting these therapies to patients who need them are discussed.
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Correction of a pathogenic gene mutation in human embryos
Hong Ma,Nuria Marti-Gutierrez,Sang-Wook Park,Jun Wu,Yeon-Mi Lee,Keiichiro Suzuki,Amy Koski,Dongmei Ji,Tomonari Hayama,Riffat Ahmed,Hayley Darby,Crystal Van Dyken,Ying Li,Eunju Kang,A. Reum Park,Daesik Kim,Sangtae Kim,Jianhui Gong,Ying Gu,Xun Xu,David Battaglia,Sacha A. Krieg,David M. Lee,Diana H. Wu,Don P. Wolf,Stephen B. Heitner,Juan Carlos Izpisua Belmonte,Paula Amato,Jin-Soo Kim,Sanjiv Kaul,Shoukhrat Mitalipov +30 more
TL;DR: The efficiency, accuracy and safety of the approach presented suggest that it has potential to be used for the correction of heritable mutations in human embryos by complementing preimplantation genetic diagnosis.
Journal ArticleDOI
Applications of CRISPR technologies in research and beyond
TL;DR: Programmable DNA cleavage using CRISPR–Cas9 enables efficient, site-specific genome engineering in single cells and whole organisms and is being used to expedite crop and livestock breeding, engineer new antimicrobials and control disease- carrying insects with gene drives.
Journal ArticleDOI
Gene therapy clinical trials worldwide to 2017: An update
Samantha L. Ginn,Anais K. Amaya,Ian E. Alexander,Ian E. Alexander,Michael Edelstein,Mohammad R. Abedi +5 more
TL;DR: This review presents the analysis of clinical trials that, to the best of the knowledge, have been or are being performed worldwide, and discusses key trends since the previous review, namely the use of chimeric antigen receptor T cells for the treatment of cancer and advancements in genome editing technologies.
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