Supplementary Figure 9: crRNA array-based transcriptional activation or knockout with AsCas12a-VPR.

(a) Schematic of the reporter cell line employed. A doxycycline-inducible TRE-RFP-shp53 reporter cell line was used. Dox withdrawal results in growth arrest and loss of RFP expression. Expression of a 15-nt TRE-guide induces RFP activation and rescues cell growth. Trp53 loss-of-function rescues cell growth. (b) Flow cytometry-based RFP quantification in mouse reporter cell line stably expressing AsCas12a-VPR transduced with different crRNA arrays harboring the 15-nt TRE-guide as indicated. Cells were cultivated in off Dox conditions. Mean ± s.d.; N = 3 independent experiments. (c) Cell growth as determined by colony formation using CV staining. N = 3 independent experiments; Representative results are shown. (d) Cell growth as determined by colony formation using crystal violet (CV). A 20-nt Trp53-guide was embedded in different crRNA arrays as indicated and lentivirally delivered to mouse cells stably expressing AsCas12a-VPR. N = 2 independent experiments; Representative results are shown. (e) Indel formation as detected by the T7EI assay. A 20-nt Trp53-guide was embedded in different crRNA arrays as indicated and lentivirally delivered to mouse cells stably expressing AsCas12a-VPR. N = 2 independent experiments; Representative results are shown.