Extended Data Fig. 3. Engineering and optimization of LwaCas13a for mammalian knockdown.
a, Knockdown of Gluc transcript by LwaCas13a and Gluc guide 1 spacers of varying length. b, Knockdown of Gluc transcript with Gluc guide 1 and varying amounts of transfected LwaCas13a plasmid. c, Knockdown of Gluc transcript by LwaCas13a and varying amounts of transfected Gluc guide 1 and 2 plasmid. d, Knockdown of Gluc transcript using guides expressed from either U6 or tRNAVal promoters. e, Knockdown of KRAS transcript using guides expressed from either U6 or tRNAVal promoters. f, Knockdown of KRAS and CXC4 transcripts by LwaCas13a using guides transfected in A375 cells with shRNA comparisons. g, Knockdown of Gluc transcript and endogenous transcripts PPIB, KRAS,and CXCR4 with active and catalytically dead Cas13a. h, Validation of the top three guides from the arrayed knockdown Gluc and Cluc screens with shRNA comparisons. i, Arrayed knockdown screen of 93 guides evenly tiled across the XIST transcript. All values are mean ± SEM with n = 3. **p< 0.01; *p< 0.05. ns = not significant. A two-tailed student's T-test was used for comparisons.