Precise in vivo genome editing via single homology arm donor mediated intron-targeting gene integration for genetic disease correction  

作　　者：Keiichiro Suzuki Mako Yamamoto Reyna Hernandez-Benitez Zhe Li Christopher Wei Rupa Devi Soligalla Emi Aizawa Fumiyuki Hatanaka Masakazu Kurita Pradeep Reddy Alejandro Ocampo Tomoaki Hishida Masahiro Sakurai Amy NNemeth Estrella Nunez Delicado Josep MCampistol Pierre Magistretti Pedro Guillen Concepcion Rodriguez Esteban Jianhui Gong Yilin Yuan Ying Gu Guang-Hui Liu Carlos Lopez-Otin Jun Wu Kun Zhang Juan Carlos Izpisua Belmonte 

机构地区：[1]Gene Expression Laboratory,Salk Institute for Biological Studies,La Jolla,CA 92037,USA [2]Institute for Advanced Co-Creation Studies,Osaka University,Osaka 560-8531,Japan [3]Graduate School of Engineering Science,Osaka University,Osaka 560-8531,Japan [4]Bioengineering,University of California,San Diego,9500 Gilman Drive,MC0412,La Jolla,CA92093-0412,USA [5]Universidad Catolica,San Antonio de Murcia,Campus de los Jeronimos,135,Guadalupe 30107,Spain [6]Hospital Clinic of Barcelona,Carrer Villarroel,170,08036 Barcelona,Spain [7]King Abdullah University of Science and Technology(KAUST),Thuwal 23955-6900,Saudi Arabia [8]Fundacion Dr.Pedro Guillen,Clinica CEMTRO,Avenida Ventisquero de la Condesa,4228035 Madrid,Spain [9]BGI-Shenzhen,Shenzhen 518083,China [10]Guangdong Provincial Key Laboratory of Genome Read and Write,Shenzhen 518120,China [11]Guangdong Provincial Academician Workstation of BGI Synthetic Genomics,BGI Shenzhen,Guangdong,China [12]Shenzhen Engineering Laboratory for Innovative Molecular Diagnostics,Shenzhen 518120,China [13]National Laboratory of Biomacromolecules,CAS Center for Excellence in Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China [14]Departamento de Bioqufmica y Biologia Molecular,Facultad de Medicina,Instituto Universitario de Oncologfa(IUOPA),Universidad de Oviedo,Oviedo,Spain [15]Department of Molecular Biology,University of Texas Southwestern Medical Center,Dallas,TX 75390,USA [16]Hamon Center for Regenerative Science and Medicine,University of Texas Southwestern Medical Center,Dallas,TX 75390,USA

出　　处：《Cell Research》2019年第10期804-819,共16页细胞研究（英文版）

基　　金：JSPS KAKENHI(15K21762 and 18H04036);Takeda Science Foundation;The Uehara Memorial Foundation;National Institutes of Natural Sciences(BS291007);The Sumitomo Foundation(170220);The Naito Foundation;The Kurata Grants(1350);Mochida Memorial Foundation;The Inamori Foundation.This research was supported by Guangdong Provincial Key Laboratory of Genome Read and Write(No.2017B030301011);Guangdong Provincial Academician Workstation of BGI Synthetic Genomics(No.2017B090904014);Shenzhen Peacock Plan(No.KQTD20150330171505310).J.C.I.B.was supported by The Leona M.and Harry B.Helmsley Charitable Trust(2012-PG-MED002);the G.Harold and Leila Y.Mathers Charitable Foundation;NIH(R01HL123755 and 5 DP1 DK113616);The Progeria Research Foundation;The Glenn Foundation,KAUST,The Moxie Foundation;Fundacion Dr.Pedro Guillen;AFE and Universidad Catolica San Antonio de Murcia(UCAM).

摘　　要：In vivo genome editing represents a powerful strategy for both understanding basic biology and treating inherited diseases.However,it remains a challenge to develop universal and efficient in vivo genome-editing tools for tissues that comprise diverse cell types in either a dividing or non-dividing state.Here,we describe a versatile in vivo gene knock-in methodology that enables the targeting of a broad range of mutations and cell types through the insertion of a minigene at an intron of the target gene locus using an intracellularly linearized single homology arm donor.As a proof-of-concept,we focused on a mouse model of prematureaging caused by a dominant point mutation,which is difficult to repair using existing in vivo genome-editing tools.Systemic treatment using our new method ameliorated aging-associated phenotypes and extended animal lifespan,thus highlighting the potential of this methodology for a broad range of in vivo genome-editing applications.

关 键 词：VIVO DONOR dividing 

分 类 号：TN9[电子电信—信息与通信工程]