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. 2021 Dec 15;50(1):549–560. doi: 10.1093/nar/gkab1191

Figure 2.

Figure 2.

Testing for orthogonality between different dCas12a-VPR variants. (A) Schematic representation of the testing of orthogonality using either native (e.g. dAsCas12a-VPR with As crRNA) or non-native (e.g. dAsCas12a-VPR with Fn crRNA) crRNA pairings. (B) The three dCas12a-VPR variants (As, Fn and Lb) were screened for orthogonality using the dual luciferase assay (as described in Figure 1B). Each dCas12a-VPR construct was delivered with each of the three targeting crRNA (As, Fn or Lb) or no targeting crRNA. Results display the mean luciferase activity when each of the three targeting crRNAs are transfected, normalised to the mean luciferase for the respective no targeting crRNA condition. Results correspond to three biological replicates, error bars show SEM and the stars (*) denote significant (P < 0.05) expression relative to no crRNA based on a Dunnett's multiple comparisons test.