Supplementary Figure 7: AsCas12a-VPR and 15-nt guides induce transcriptional activation in vivo.
From: Multiplexed orthogonal genome editing and transcriptional activation by Cas12a
(a) HDTV of AsCas12a-VPR, crRNA expressing vectors, and SB into TRE-GFP reporter knock-in mice. crRNAs with a 20-nt Rosa26 ctrl-guide, as well as a 20-nt TRE-guide and the corresponding 15-nt TRE-guide were tested. 2 weeks post-HDTV, expression of GFP in livers was investigated by immunofluorescence. BFP staining indicates crRNA expression. DAPI staining visualizes nuclei. N = 3 mice per group. Representative images are shown. Scale bar = 100 µm. (b) HDTV of AsCas12a-VPR, 15-nt Krt19-guide expressing vectors, and SB into mice. Immunohistochemistry of Krt19. crRNAs with a 15-nt Rosa26 ctrl-guide, as well as 15-nt KRT19-guides were tested. Livers were analyzed 2 weeks post-HDTV. * = Krt19 positive bile duct cells/cholangiocytes; arrows = Krt19 positive hepatocytes. N = 5 animals per group. Representative results are shown. Scale bar = 20 µm.
