Figure 2. CRISPRa screens reveal metabolic genes conferring resistance to iron depletion.

(A) BafA1 and DFO decreases cellular iron availability through different mechanisms.

(B) Dose-dependent effects of DFO and BafA1 on proliferation of Jurkat cells (Mean ± SEM, n = 5).

(C) Gene scores of untreated vs. DFO (2 μM)-treated Jurkat-dCas9a cells (left). Gene score is the median log₂ fold change in the abundance of all sgRNAs targeting that gene during the screening period. Differential gene scores of top 20 genes providing resistance to DFO (right). Iron/heme-related genes are indicated in dark blue.

(D) Gene scores of untreated vs. BafA1 (5 nM)-treated Jurkat-dCas9a cells (left). Differential gene scores of top 20 genes providing resistance to BafA1 (right). Vacuolar ATPase subunits are indicated in light blue, lipid metabolism-related genes in green, other genes in gray.

(E) Scheme displaying selected scoring genes conferring resistance to iron-restriction.

(F) Immunoblot analysis of control and SLC25A37-overexpressing Jurkat cells. β-actin was used as loading control.

(G) Fold change in the number (log₂) of control vector and SLC25A37-overexpressing cells after 5 days in the presence of indicated DFO concentrations (left). Representative bright-field micrographs of Jurkat cells after 5-day treatment with 3 μM DFO (right). Scale bar is 50 μM.

See also Figures S1-S3, Tables S9-S14