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Title: CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae
Authors: Li, Zhen-Hai
Liu, Min
Lyu, Xiao-Mei
Wang, Feng-Qing
Wei, Dong-Zhi
Keywords: Engineering::Bioengineering
Issue Date: 2018
Source: Li, Z.-H., Liu, M., Lyu, X.-M., Wang, F.-Q., & Wei, D.-Z. (2018). CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae. Journal of Basic Microbiology, 58(12), 1100-1104. doi:10.1002/jobm.201800195
Journal: Journal of Basic Microbiology
Abstract: In this study, we focused on the applicability of CRISPR/Cpf1 in genome simplification of Saccharomyces cerevisiae and established a CRISPR/Cpf1 assisted method for rapid markerless large fragment deletion to facilitate laboratory evolution of geome of S. cerevisiae by rational genetic engineering. This method uses a Cpf1 expression plasmid and a crRNA array expression plasmid. The DNA fragment between two DSBs generated by CRISPR/Cpf1 can be cut off from the chromosome, along with re-ligation of the genomic endpoints of the DSBs. Using this method, the large DNA fragment of ∼38 kb between the two genes of TRM10 and REX4 was successfully and rapidly deleted, which was verified by PCR and Sanger DNA Sequencing. This method is simple and rapid, and can be easily implemented for large fragment deletion in S. cerevisiae.
ISSN: 0233-111X
DOI: 10.1002/jobm.201800195
Rights: © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. All rights reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:SCBE Journal Articles

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