Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ
Name
s12864-016-3331-9.pdf
Size
4 MB
Format
Adobe PDF
Checksum (MD5)
09f68109312a501b2fefdc3dd79a85b1
Author(s)
Aida, Tomomi
Nakade, Shota
Sakuma, Tetsushi
Izu, Yayoi
Oishi, Ayu
Mochida, Keiji
Ishikubo, Harumi
Usami, Takako
Aizawa, Hidenori
Yamamoto, Takashi
Date Issued
November 2016
Journal
BMC Genomics
Publisher
Biomed Central Ltd
Citation
Aida, Tomomi et al. "Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ." BMC Genomics 17 (November 2016): 979 © 2016 The Author(s)
Version
Final published version
Abstract
Background: Although CRISPR/Cas enables one-step gene cassette knock-in, assembling targeting vectors containing long homology arms is a laborious process for high-throughput knock-in. We recently developed the CRISPR/Cas-based precise integration into the target chromosome (PITCh) system for a gene cassette knock-in without long homology arms mediated by microhomology-mediated end-joining. Results: Here, we identified exonuclease 1 (Exo1) as an enhancer for PITCh in human cells. By combining the Exo1 and PITCh-directed donor vectors, we achieved convenient one-step knock-in of gene cassettes and floxed allele both in human cells and mouse zygotes. Conclusions: Our results provide a technical platform for high-throughput knock-in.
MIT Department
McGovern Institute for Brain Research at MIT
Terms of Use
Creative Commons Attribution 4.0 International
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1186/s12864-016-3331-9
