Fig. 3: Cas13d orthologs tolerate multiple domain deletions while maintaining target-binding and RNA-knockdown activity.
From: A strategy for Cas13 miniaturization based on the structure and AlphaFold
a Schematic diagram showing the miniaturization of EsCas13d to Cas13d orthologs using the IDC strategy and AlphaFold2. b Comparison of the NTD domains of RfxCas13d and mini-RfxCas13d, EsCas13d and mini-EsCas13d, RspCas13d and mini-RspCas13d. RfxCas13d, mini-RfxCas13d, RspCas13d, mini-RspCas13d and mini-EsCas13d were predicted by AlphaFold2, except for EsCas13d (6E9E). Deletion regions were colored deep gray. c Schematic showing the domain organization of EsCas13d and mini-EsCas13d. d Schematic showing the domain organization of RspCas13d and mini-RspCas13d. e Comparison of the EsCas13d and mini-EsCas13d knockdown efficiency of endogenous transcripts. The data are presented as the mean ± SD. Two-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons. *P < 0.05, **P < 0.01. (n = 3 biological replicates, each with an average of 4 technical replicates). Source data are provided as a Source data file. f Comparison of the endogenous transcripts knockdown efficiency of RspCas13d and mini-RspCas13d. NT non-target crRNA. The data are presented as the mean ± SD. Two-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons. *P < 0.05, **P < 0.01, ns, not significant. (n = 3 biological replicates, each with an average of 4 technical replicates). Source data are provided as a Source data file.
