Journal: Cell reports

Article Title: Biphasic control of the B cell transcriptome by mTORC1 and GSK3

doi: 10.1016/j.celrep.2025.116361

Figure Lengend Snippet: (A) Schematic workflow of the CRISPR screen approach. OCAB reporter cells are created through mOrange-P2A knock-in, transduced with the genome-wide sgRNA library Brunello, and FACS-sorted to identify the activators and repressors of OCAB expression. (B) Distribution of OCAB activators ranked by the RRA score. Selected activators were individually validated with specific targeting sgRNAs. A total of 113 hits were tested with 74 true positives. (C) mO-OCAB expression measured by flow cytometry 10 days after FLI1 or control sgRNA transduction. (D) ChIP-seq (FLI1, BRD4) and RNA-seq profiles at the OCAB locus in control and FLI1-depleted cells. Fold change and adjusted p values (Benjamini-Hochberg) are indicated. (E) As in (C) but after SPEN or control sgRNA transduction.

Article Snippet: For this, selected sgRNAs and a non-targeting sgRNA from the Brunello library were cloned into lentiGuide-puro (Addgene).

Techniques: CRISPR, Knock-In, Transduction, Genome Wide, Expressing, Flow Cytometry, Control, ChIP-sequencing, RNA Sequencing