Comparative optimization of combinatorial CRISPR screens
Abstract
Combinatorial CRISPR technologies have emerged as a transformative approach to systematically probe genetic interactions and dependencies of redundant gene pairs. However, the performance of different functional genomic tools for multiplexing sgRNAs vary widely. Here, we generate and benchmark ten distinct pooled combinatorial CRISPR libraries targeting paralog pairs to optimize digenic knockout screens. Libraries composed of dual Streptococcus pyogenes Cas9 (spCas9), orthogonal spCas9 and Staphylococcus aureus (saCas9), and enhanced Cas12a from Acidaminococcus were evaluated. We demonstrate a combination of alternative tracrRNA sequences from spCas9 consistently show superior effect size and positional balance between the sgRNAs as a robust combinatorial approach to profile genetic interactions of multiple genes.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- May 05, 2022
- Source ID
- 10.1038/s41467-022-30196-9
Entities
People
- Christopher R. Vakoc
- Davide Monducci
- Devishi Kesar
- Diego J. Rodriguez
- Joshua M Dempster
- Mahdi Zamanighomi
- Michael J. Young
- Olaf Klingbeil
- Ruitong Li
- Takahiro Ito
- William R Sellers
- Xiaoping Yang
- Zaid Bayyat
Organizations
- Ludwig Institute for Cancer Research
- United States Department of Defense