(d)Cas12a-anti-CRISPR circuits

Testing the usage of both dCas12a-based transcription factors and AcrVAs as components for synthetic gene circuits in the yeast S. cerevisiae.
Conclusion:
In the baker’s yeast, two dCas12a proteins (denAsCas12a and dLbCas12a) work as both activators (upon fusion to a strong activation domain) and repressors, whereas dMbCa12a (where the first AcrVAs were found) is unfunctional. Moreover, AcrVA1, AcrVA4, and AcrVA5 are able to inhibit denAsCas12a (AcrVA1 only) and dLbCa12a. AcrVA4 could also significantly limit gene editing by LbCas12a. dCas12a:crRNA and AcrVA proteins are highly-performant components in S. cerevisiae synthetic transcriptional networks.
Notes:
1-1: 1x1-pGAL1-dMb-VPR-pSNR53i-bA 1-2: 1x1-pGAL1-dMb-VPR-pSNR53i-bS 1-3: 1x1-pGAL1-dMb-VPR-pSNR53i-Sc-crRNA 2-1: 3x1-pGAL1-dMb-VPR-pSNR53i-bA 2-2: 3x1-pGAL1-dMb-VPR-pSNR53i-bS 2-3: 3x1-pGAL1-dMb-VPR-pSNR53i-Sc-crRNA 3-1: 1x1-pGAL1-dMb-VP64-pSNR53i-bA 3-2: 1x1-pGAL1-dMb-VP64-pSNR53i-bS 3-3: 1x1-pGAL1-dMb-VP64-pSNR53i-Sc-crRNA 4-1: 3x1-pGAL1-dMb-VP64-pSNR53i-bA 4-2: 3x1-pGAL1-dMb-VP64-pSNR53i-bS 4-3: 3x1-pGAL1-dMb-VP64-pSNR53i-Sc-crRNA 5-1: 1x1-pGPD-dMb-VPR-pSNR53i-bA 5-2: 1x1-pGPD-dMb-VPR-pSNR53i-bS 5-3: 1x1-pGPD-dMb-VPR-pSNR53i-Sc-crRNA 6-1: 3x1-pGPD-dMb-VPR-pSNR53i-bA 6-2: 3x1-pGPD-dMb-VPR-pSNR53i-bS 6-3: 3x1-pGPD-dMb-VPR-pSNR53i-Sc-crRNA 7-1: 1x1-pGPD-dMb-VP64-pSNR53i-bA 7-2: 1x1-pGPD-dMb-VP64-pSNR53i-bA 7-3: 1x1-pGPD-dMb-VP64-pSNR53i-Sc-crRNA 8-1: 3x1-pGPD-dMb-VP64-pSNR53i-bA 8-2: 3x1-pGPD-dMb-VP64-pSNR53i-bS 8-3: 3x1-pGPD-dMb-VP64-pSNR53i-Sc-crRNA