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

Keiichiro SuzukiMako YamamotoReyna Hernandez-BenitezZhe LiChristopher WeiRupa Devi SoligallaEmi AizawaFumiyuki HatanakaMasakazu KuritaPradeep ReddyAlejandro OcampoTomoaki HishidaMasahiro SakuraiAmy N NemethEstrella Nuñez DelicadoJosep M CampistolPierre MagistrettiPedro GuillenConcepcion Rodriguez EstebanJianhui GongYilin YuanYing GuGuang-Hui LiuCarlos López-OtínJun WuKun ZhangJuan Carlos Izpisua Belmonte
Published in: Cell research (2019)
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 premature-aging 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.
Keyphrases
  • genome editing
  • crispr cas
  • genome wide
  • copy number
  • single cell
  • genome wide identification
  • gene expression
  • dna methylation
  • transcription factor
  • replacement therapy
  • climate change