blasticidin-selectable lentiviral cassette allowing stable expression of the cas9 nuclease  (Addgene inc)

Journal: bioRxiv

Article Title: The Role of ATP Synthase Subunit e (ATP5I) in Mediating the Metabolic and Antiproliferative Effects of Biguanides

doi: 10.1101/2024.09.20.614047

Figure Lengend Snippet: A) Results of the pooled genome-wide CRISPR/Cas9 KO screen made in NALM-6 cells treated with 16 mM metformin or control. Data are represented as a Volcano plot of gene enrichment/depletion scores vs p-values from using the CRANKS algorithm. Some genes of interest are labeled. Enhancers of metformin growth inhibition with negative CRANKS scores below 2.5 (dashed line) are labeled blue, while suppressors with positive CRANKS scores above 2.5 (dashed line) are labeled red. B-C) Pairwise comparison of gene CRANKS scores obtained from screening metformin 16 mM in NALM-6 cells against that from screening either 70 nM rotenone or (C) 2 μM oligomycin A. D) Model for metformin action triggering the OMA1-DELE1-HRI pathwat.

Article Snippet: Briefly, a NALM-6 clone bearing an integrated doxycycline-inducible Cas9 expression cassette generated by lentiviruses made from pCW-Cas9 (Addgene #50661) was transduced with the genome-wide KO EKO sgRNA library (278,754 different sgRNAs).

Techniques: Genome Wide, CRISPR, Control, Labeling, Inhibition, Comparison