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© Research
Publication : Nature biotechnology

Dual gene activation and knockout screen reveals directional dependencies in genetic networks

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nature biotechnology - 15 Jan 2018

Boettcher M, Tian R, Blau JA, Markegard E, Wagner RT, Wu D, Mo X, Biton A, Zaitlen N, Fu H, McCormick F, Kampmann M, McManus MT

Link to Pubmed [PMID] – 29334369

Nat. Biotechnol. 2018 Feb;36(2):170-178

Understanding the direction of information flow is essential for characterizing how genetic networks affect phenotypes. However, methods to find genetic interactions largely fail to reveal directional dependencies. We combine two orthogonal Cas9 proteins from Streptococcus pyogenes and Staphylococcus aureus to carry out a dual screen in which one gene is activated while a second gene is deleted in the same cell. We analyze the quantitative effects of activation and knockout to calculate genetic interaction and directionality scores for each gene pair. Based on the results from over 100,000 perturbed gene pairs, we reconstruct a directional dependency network for human K562 leukemia cells and demonstrate how our approach allows the determination of directionality in activating genetic interactions. Our interaction network connects previously uncharacterized genes to well-studied pathways and identifies targets relevant for therapeutic intervention.

https://www.ncbi.nlm.nih.gov/pubmed/29334369