@inbook{2b1146013e2f48558e889d367dbf92dd,
title = "Delta integration CRISPR-Cas (Di-CRISPR) in saccharomyces cerevisiae",
abstract = "Despite the advances made in genetic engineering of Saccharomyces cerevisiae, the multicopy genomic integration of large biochemical pathways remains a challenge. Here, we developed a Di-CRISPR (delta integration CRISPR-Cas) platform based on cleavage of the delta sites by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated systems (Cas) to enable unprecedented high-efficiency, multicopy, markerless integrations of large biochemical pathways into the S. cerevisiae genome. Detailed protocols are provided on the entire workflow which includes pDi-CRISPR plasmid and donor DNA construction, Di-CRISPR-mediated integration and analysis of integration efficiencies and copy numbers through flow cytometry and quantitative polymerase chain reaction (qPCR).",
keywords = "CRISPR-Cas, Delta integration, Genome engineering, Genome integration, Saccharomyces cerevisiae, Synthetic biology",
author = "Shuobo Shi and Youyun Liang and Ang, {Ee Lui} and Huimin Zhao",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2019.",
year = "2019",
doi = "10.1007/978-1-4939-9142-6_6",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "73--91",
booktitle = "Methods in Molecular Biology",
}