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

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

Cell reports (Cambridge), 2022-01, Vol.38 (1), p.110196-110196, Article 110196 [Periódico revisado por pares]

United States

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Título:
Robust genome editing in adult vascular endothelium by nanoparticle delivery of CRISPR-Cas9 plasmid DNA
Autor: Zhang, Xianming ; Jin, Hua ; Huang, Xiaojia ; Chaurasiya, Birendra ; Dong, Daoyin ; Shanley, Thomas P ; Zhao, You-Yang
Assuntos: Animals ; Cell Line ; Clustered Regularly Interspaced Short Palindromic Repeats - genetics ; CRISPR-Associated Protein 9 - genetics ; CRISPR-Cas Systems - genetics ; Endothelium, Vascular - cytology ; Female ; Gene Editing - methods ; Genetic Therapy - methods ; High-Throughput Nucleotide Sequencing ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Nanoparticles - chemistry ; Plasmids - genetics ; Polyethyleneimine - chemistry ; RNA, Guide, CRISPR-Cas Systems - genetics
É parte de: Cell reports (Cambridge), 2022-01, Vol.38 (1), p.110196-110196, Article 110196
Notas: X.Z., H.J., X.H., T.P.S., and Y.Y.Z. conceptualized the study; X.Z., H.J., X.H., B.C., D.D., and Y.Y.Z. designed and performed the experiments; X.Z., X.H., and Y.Y.Z. wrote the manuscript with input from all authors.
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Descrição: 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.
Editor: United States
Idioma: Inglês

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

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

United States

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