Fig. 6.

S10 peptide delivery of Cas12a RNP shows editing in ROSA^mT/mG locus in vivo. a Schematic of Cas12a gRNA targeting both loxP sequences flanking tdTomato gene in ROSA^mT/mG mice. S10 peptide and Cas12a RNP (2.5 µM Cas12a; 2.0 µM gRNA) directed to suboptimal PAM target in loxP sites flanking the tdTomato cassette administered twice a day for 2 days, via intranasal instillation. Seven days later, conversion of tdTomato to GFP expression was visualized in lung tissue sections. b Cas12a mediated deletion of tdTomato in ROSA^mT/mG in large airway epithelia in vivo; ×2 magnification. c Editing in a large airway; ×20 magnification. d Editing in a small airway; ×20 magnification. e Co-localization of GFP and marker of ciliated cells (α-tubulin, white) in large airway. Arrowheads indicate GFP and α-tubulin co-localization. Arrow denotes edited (GFP⁺) non-ciliated cell negative for α-tubulin; ×40 magnification. f Distal lung region. Co-localization of GFP⁺ and SP-C (white) identifies alveolar type II cells (arrowhead); ×40 magnification. g Editing efficiency of Cas12a RNP in large and small airways quantified by the number of GFP⁺ cells. Horizontal lines indicate mean ± SE; n = 3 mice/group. Data underlying this figure are provided as Source Data file