Figure 3.

Target RNA cleavage of Cas13 variants in silkworm BmE cells

A–D’: Microscopy analysis of changes in green fluorescence in EGFP-Cas13-expressed BmE cells following transfection of crRNAs targeting EGFP. White light microscopy (A–D) and fluorescence microscopy (A’–D’) were conducted. Scale bar: 50 μm. E, F: RT-qPCR analysis of EGFP transcripts in EGFP-Cas13-expressed BmE cells following transfection of EGFP-targeting crRNAs. RpL3 and Actin genes were used as an internal control. Significant differences were determined using Student’s t-test. ^*: P<0.05; ^**: P<0.01; ^***: P<0.001 versus control. G, H: Western blotting validation of EGFP protein expression changes in EGFP-Cas13-expressed BmE cells following transfection with EGFP-targeting crRNAs. Anti-EGFP and anti-Tubulin antibodies were used. Control, transfection of crRNA targeting DsRed transcript; EGFP-i, RNA cleavage following transfection of EGFP-targeting crRNA.