Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ. (December 2016)
- Record Type:
- Journal Article
- Title:
- Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ. (December 2016)
- Main Title:
- Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ
- Authors:
- Aida, Tomomi
Nakade, Shota
Sakuma, Tetsushi
Izu, Yayoi
Oishi, Ayu
Mochida, Keiji
Ishikubo, Harumi
Usami, Takako
Aizawa, Hidenori
Yamamoto, Takashi
Tanaka, Kohichi - Abstract:
- 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 identifiedexonuclease 1 (Exo1 ) as an enhancer for PITCh in human cells. By combining theExo1 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.
- Is Part Of:
- BMC genomics. Volume 17:Number 1(2016)
- Journal:
- BMC genomics
- Issue:
- Volume 17:Number 1(2016)
- Issue Display:
- Volume 17, Issue 1 (2016)
- Year:
- 2016
- Volume:
- 17
- Issue:
- 1
- Issue Sort Value:
- 2016-0017-0001-0000
- Page Start:
- 1
- Page End:
- 18
- Publication Date:
- 2016-12
- Subjects:
- MMEJ -- CRISPR/Cas -- Gene cassette -- Reporter -- Flox -- Knock-in -- Mouse -- Exo1 -- High throughput -- Cloning-free
Genomes -- Periodicals
Gene mapping -- Periodicals
Genomics -- Periodicals
Base Sequence -- Periodicals
Chromosome Mapping -- Periodicals
Genetic Techniques -- Periodicals
Sequence Analysis, DNA -- Periodicals
572.8605 - Journal URLs:
- http://www.biomedcentral.com/bmcgenomics/ ↗
http://www.pubmedcentral.nih.gov/tocrender.fcgi?journal=32 ↗
http://link.springer.com/ ↗ - DOI:
- 10.1186/s12864-016-3331-9 ↗
- Languages:
- English
- ISSNs:
- 1471-2164
- Deposit Type:
- Legaldeposit
- View Content:
- Available online (eLD content is only available in our Reading Rooms) ↗
- Physical Locations:
- British Library DSC - BLDSS-3PM
British Library STI - ELD Digital store - Ingest File:
- 9960.xml