Robust genome editing in adult vascular endothelium by nanoparticle delivery of CRISPR-Cas9 plasmid DNA

Xianming Zhang, Hua Jin, Xiaojia Huang, Birendra Chaurasiya, Daoyin Dong, Thomas P. Shanley, You Yang Zhao*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Vascular endothelium plays a crucial role in vascular homeostasis and tissue fluid balance. To target endothelium for robust genome editing, we developed poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolide) (PEG-b-PLGA) copolymer-based nanoparticle formulated with polyethyleneimine. A single i.v. administration of mixture of nanoparticles and plasmid DNA expressing Cas9 controlled by CDH5 promoter and guide RNA (U6 promoter) induced highly efficient genome editing in endothelial cells (ECs) of the vasculatures, including lung, heart, aorta, and peripheral vessels in adult mice. Western blotting and immunofluorescent staining demonstrated an ∼80% decrease of protein expression selectively in ECs, resulting in a phenotype similar to that of genetic knockout mice. Nanoparticle delivery of plasmid DNA could induce genome editing of two genes or genome editing and transgene expression in ECs simultaneously. Thus, nanoparticle delivery of plasmid DNA is a powerful tool to rapidly and efficiently alter expression of gene(s) in ECs for cardiovascular research and potential gene therapy.

Original languageEnglish (US)
Article number110196
JournalCell reports
Volume38
Issue number1
DOIs
StatePublished - Jan 4 2022
Externally publishedYes

Keywords

  • CRISPR-Cas9
  • cardiovascular diseases
  • endothelial cells
  • endothelium targeting
  • gene delivery
  • gene therapy
  • genome editing
  • lung diseases
  • nanoparticle
  • non-viral CRISPR delivery

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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