Fig. 1 Intra-amniotic delivery of CRISPR-Cas9 results in pulmonary gene editing. Intra-amniotic delivery of CRISPR-Cas9 results in pulmonary gene editing.

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Fig. 1 Intra-amniotic delivery of CRISPR-Cas9 results in pulmonary gene editing. Intra-amniotic delivery of CRISPR-Cas9 results in pulmonary gene editing. (A) Schematic representation of intra-amniotic route of fetal lung gene editing. (B) Experimental design of gene editing in R26mTmG/+ mice. (C) Fluorescent stereomicroscopy, using a filter to detect tdTomato and EGFP, of lungs from R26mTmG/+ mice injected with Ad.Cre, Ad.mTmG, or Ad.Null. (D) IHC for EGFP and tdTomato expression in the proximal airway and distal air saccules of lungs from R26mTmG/+ mice injected with Ad.Cre, Ad.mTmG, or Ad.Null. White arrowheads indicate EGFP staining. (E) PCR assay using primers to detect the on-target editing in DNA isolated from E19 lungs of R26mTmG/+ mice injected with Ad.Cre, Ad.mTmG, or Ad.Null. Edited band, 545 base pairs (bp); unedited band, 2951 bp. n = 2 to 6 per group. One fetus that was injected with Ad.mTmG and lacked notable EGFP fluorescence (GFP−) was also negative for gene editing by PCR, indicating a likely technical failure at the time of injection. (F) Sanger sequencing of the 545-bp edited mTmG PCR product from an R26mTmG/+ mouse injected with Ad.mTmG. (G) Sanger sequencing of the 545-bp Cre-recombined mTmG PCR product from an R26mTmG/+ mouse injected with Ad.Cre. Scale bars, 1000 μm (C) and 50 μm (D). IA, intra-amniotic; E, gestational day. Deepthi Alapati et al., Sci Transl Med 2019;11:eaav8375 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works