Extended Data Fig. 8: Further characterization of RNA knockdown in mammalian cells by Cas7-11.

a, Knockdown of Gluc mRNA in mammalian cells by DiCas7-11 with a panel of guides tiled across the transcript. b, Knockdown of Gluc mRNA in mammalian cells by DiCas7-11 with N- and C-terminus fusions of nuclear export signal (NES) tags (Adeno type 5 E1B-55K) with guides tiled across the transcript. c, Knockdown activity in HEK293FT cells of DiCas7-11, shRNA, LwaCas13a, PspCas13b, and RfxCas13d against the Gluc transcript normalized to corresponding non-targeting controls. All guides are designed to target the same region of the Gluc transcript. d, Comparison of knockdown activity of Gluc mRNA in mammalian cells between active DiCas7-11, catalytically inactive D429A/D654A DiCas7-11, and GFP. Guides have the full DR sequence. e, Knockdown activity of DiCas7-11 against the multiple endogenous transcripts normalized to two non-targeting conditions. Dotted lines represent background expression and 65% knockdown by DiCas7-11. f, Knockdown of EGFP and mCherry mRNA in mammalian cells by DiCas7-11 with a panel of tiling guides. g, Knockdown of Gluc mRNA in mammalian cells by DiCas7-11 with a panel of guides of different lengths. h, Knockdown of Gluc mRNA in mammalian cells by hvsCas7-11 with a panel of guides containing the full DR sequence. Guides are designed to be tiled across the Gluc transcript. i, Knockdown of Gluc mRNA in HepG2 cells with hvsCas7-11 and no protein conditions using guides 1 and 11 as well as a non-targeting guide. j, Knockdown of Gluc mRNA in HEK293FT cells with hvsCas7-11 modified with an NES sequence (Adeno type 5 E1B-55K) and using Gluc guide 1. The NES tag is either C-terminally fused or fused on both ends of the hvsCas7-11 protein. All data in this figure are mean ± s.e.m.; n = 2 or n = 3, as shown.