Fig. 4: Application of the dCas13b-tethering systems for cell-based validation and mechanistic studies.
From: IGF2BP3 promotes mRNA degradation through internal m7G modification
a Schematic diagram showing the modified dCas13b system with IGF2BP3 (dCas13b-IGF2BP3, left) and METTL1 (dCas13b-METTL1, right). b Relative m7G methylation levels of the selected gene loci. Mean ± SEM of three independent experiments. c Changes in mRNA levels in HepG2 cells with introduction of dCas13b-IGF2BP3 and guide RNA at the loci or not (neg). Mean ± SEM (n = 3) with two-tailed Student’s t-tests. Calculated half lifetimes are marked in the corresponding colors. d Relative m7G methylation level of the loci of the selected genes upon introducing the dCas13b-METTL1 tethering with the guide RNAs at the target loci or not (neg), normalized to the methylation level in the wildtype cells. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. P-values are marked at the top of each group. e Changes in mRNA levels in HepG2 cells with the introduction of dCas13b-METTL1 and the guide RNAs at the loci or not (neg). Mean ± SEM (n = 3) with two-tailed Student’s t-tests. Calculated half lifetimes are marked in the corresponding colors. f Western blotting of EXOSC2 in cell lysate from in vitro pulldown in HepG2 cells using the antibodies that recognize IGF2BP1, IGF2BP2, or IGF2BP3. Rabbit IgG was used as a negative control. This experiment was repeated independently twice with similar results. g mRNA levels of the m7G targets upon IGF2BP3 overexpression, or EXOSC2 knockdown, or both. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. All source data are provided as a Source Data file.
