Efficient single-copy HDR by 5' modified long dsDNA donors

ELife
Jose Arturo Gutierrez-TrianaJoachim Wittbrodt

Abstract

CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5' ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful gene replacement or tagging.

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Citations

Mar 6, 2019·PloS One·Omar T HammoudaThomas Thumberger
Jul 10, 2019·ELife·Leon Hilgers, Julia Schwarzer
Feb 9, 2020·Nature Biotechnology·Alexis L NorrisHeather A Lombardi
May 15, 2020·The Journal of Cell Biology·Julia FuellerMichael Knop
Aug 23, 2020·International Journal of Molecular Sciences·Zhihua ZhaoJianzhong Jeff Xi
Feb 20, 2019·Molecular Genetics and Genomics : MGG·Robert HerzogFlorian Hennicke
Aug 29, 2019·International Journal of Molecular Sciences·Baubak BajoghliNarges Aghaallaei
Jul 30, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Joshua BushRemi Veneziano
Feb 18, 2020·Nucleic Acids Research·Sara E DiNapoliYariv Houvras
Nov 7, 2020·Development, Growth & Differentiation·Yu MurakamiMasato Kinoshita
Jun 10, 2021·Development·Sevinç GücümJoachim Wittbrodt
Aug 28, 2021·International Journal of Molecular Sciences·Christopher E DenesDaniel Hesselson
Jan 25, 2022·Genesis : the Journal of Genetics and Development·Yu Murakami, Toru Kobayashi

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Methods Mentioned

BETA
PCR
genotyping
electrophoresis
transgenic
in vitro transcription

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