Related Experiment Video
Updated: Aug 5, 2026

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
Published on: May 28, 2019
Protocol for CRISPR genome editing in S. cerevisiae using PCR-based guide insertion
Hosein Rostamian1, Ethan W Madden1, Frank M Kaplan2
1Department of Biochemistry & Biophysics, The University of North Carolina School of Medicine, Chapel Hill, NC, USA; Curriculum in Genetics and Molecular Biology, The University of North Carolina School of Medicine, Chapel Hill, NC, USA.
Abstract:
Here, we present a CRISPR-Cas9 genome-editing protocol for Saccharomyces cerevisiae that simplifies guide construction and enables rapid guide iteration by replacing restriction/ligation cloning with PCR-based guide installation and seamless Cas9-plasmid recircularization. We describe single-guide RNA (sgRNA) and homology-directed repair (HDR) donor design, guide insertion into a KanMX/G418-selectable Cas9-sgRNA plasmid, plasmid cloning and sequence verification in E. coli, and lithium acetate/polyethylene glycol (LiAc/PEG) co-transformation of yeast with the verified plasmid and HDR donor.
Related Concept Videos
CRISPR/Cas9 Genome Editing
CRISPR

