AAV-CRISPR Somatic Editing

Overview

AAV-CRISPR somatic editing is a platform for in vivo somatic genome engineering using adeno-associated virus (AAV) vectors to deliver CRISPR-Cas9 guide RNAs (and optionally HDR donor templates) to target tissues in Cas9-transgenic animals. In the mammary gland context, AAV serotype 2/9 is delivered intraductally to achieve luminal-epithelial-specific editing of cancer-relevant genes (e.g., Tp53, Nf1, Pik3ca, Kras) either as frameshift-inducing Indels or precise homology-directed repair (HDR) substitutions. The approach enables rapid, genotype-controlled tumor modeling in immunocompetent hosts without germline transgenics.

Used by

  • Intraductal AAV-CRISPR delivery into Cas9-transgenic rats was used to engineer six distinct mammary tumor genotypes (Tp53Indel, Nf1Indel, Nf1Indel/Tp53Indel, Pik3caH1047R, Pik3caH1047R/Tp53Indel, triple-edit) modeling ER+/PR+ ductal carcinoma; single-gene Nf1 editing produced DCIS at 4-week median latency, triple editing (AAV-PNT) at 8-day median latency. PMID:26437033
  • LentiCRISPR v2 nickase (D10A Cas9) was used to knock out CDH1 in MGHU4 and RT4 bladder cancer cell lines; knockout enhanced cell migration in wound-healing scratch assay and Boyden chamber assay (p<0.05), providing mechanistic evidence for the plasmacytoid variant’s peritoneal spread pattern. PMID:26901067

Notes

  • Editing efficiency in rat mammary epithelium: ~4.2% Indel rate in whole-gland alleles (day 4), estimated 10.5–21% in luminal epithelial cells; HDR-based Pik3caH1047R achieved ~40% allelic editing in tumors.
  • Key advantage over GEM models: flexible combinatorial driver testing in an immunocompetent host without the need to maintain complex germline crosses.
  • Species context matters: identical edits in mice produce ER-/PR- metaplastic tumors, whereas rats develop ER+/PR+ ductal carcinoma.
  • AAV serotype 2/9 used for intraductal delivery; titer typically 2×10^10 – 2.8×10^11 gc/gland depending on construct.

Sources

This page was processed by crosslinker on 2026-05-14. - PMID:26901067

This page was processed by entity-page-writer on 2026-05-15.