Fig. 4: Multiplexed base editing by Cas12a variant-mediated base editors in embryos.
From: Multiplexed base editing through Cas12a variant-mediated cytosine and adenine base editors
a Construct of CRISPR array of pcr4 with four tandem sgRNAs targeting to porcine genome. The sequences of individual targets are listed below. PAMs are marked in purple and target Cs are marked in red. b Schematic illustrating the workflow of RNA mixture microinjection of porcine parthenogenetic (PA) embryos to assess the editing efficiency. c Simultaneous C-to-T editing of four loci (PUM1, GHR, HMGA2, and PUM2) by RR-A3A-Y130F with multiplex-sgRNA (CRISPR array pcr4) or pooled single sgRNAs in porcine PA embryos (n = 11). d Efficiencies of C-to-T editing of target Cs (marked in red in a) induced by RR-A3A-Y130F at four indicated loci (n = 11). e Representative images of porcine PA embryos development in vitro six days after RNA injection, with uninjected PA embryos as the control (n = 4). Scale bar, 200 µm. f Development of porcine PA embryos in vitro after RNA injection (n = 4). Values and error bars for c and f represent the mean and SEM, respectively. Values for d represent the median with interquartile range. Statistical significance was calculated by unpaired two-tailed Student’s t-test (*p < 0.05).
