TY  - GEN
T1  - Highly improved cloning efficiency for plasmid-based CRISPR knock-in in C. elegans
AU  - DeMott, Ella
AU  - Dickinson, Daniel J
AU  - Doonan, Ryan
DO  - 10.17912/micropub.biology.000499
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000499/
AB  - The Glow Worms research stream is a course-based undergraduate research education (CURE) laboratory that focuses on making fluorescent protein knock-in strains at genetic loci of interest to the research community. To facilitate making strains, we aimed to determine whether the self-excising cassette (SEC) selection method for CRISPR knock-in could be improved. Indeed, we recently reported that using high-quality DNA and reducing Cas9/sgRNA concentration facilitates recovery of homology directed repair (HDR) knock-ins, requiring microinjection of only 10 P0 worms (Huang et al. 2021). Here, we show that cloning of SEC-based CRISPR plasmids can also be highly efficient and reproducible when optimized based on quantitative considerations.
PY  - 2021
JO  - microPublication Biology
ER  -