Figure 1. ERK Activation Promotes mRNA m⁶A Methylation.

(A) Schematic diagram of a circular RNA (circRNA) translation reporter consisting of a single exon and two introns with complementary sequences. The exon containing GGACU can be back-spliced to generate circRNAs that drive GFP translation.

(B) Overview of the CRISPR screen. Cas9 KO libraries are packaged into lentivirus and then transduced into HeLa cells contain circRNA GFP reporters. Cells with the top and bottom 5% GFP expression were collected by flow cytometry. The sgRNA were amplified from genomic DNA and then sequenced, followed by statistical analyses to identify candidate genes.

(C) Positive regulators for the m⁶A pathway identified in the screen using circular GGACU-GFP reporters.

(D) Pathway analysis of sgRNA enriched in the bottom 5% GFP cells with circRNA GFP reporters.

See also Figure S1, Table S1, and Table S2.