LNP-mediated delivery of CRISPR RNP for wide-spread in vivo genome editing in mouse cornea

被引：37

作者：

Mohanna, Seyedeh Zeinab Mirjalili ^{[1
,2
]}

Djaksigulova, Diana ^{[1
]}

Hill, Austin M. ^{[3
]}

Wagner, Pamela K. ^{[3
]}

Simpson, Elizabeth M. ^{[1
,2
,4
]}

Leavitt, Blair R. ^{[1
,2
,3
]}

机构：

[1] Univ British Columbia, British Columbia Childrens Hosp, Ctr Mol Med & Therapeut, Vancouver, BC, Canada

[2] Univ British Columbia, Dept Med Genet, Vancouver, BC, Canada

[3] Incis Genet Inc, Vancouver, BC, Canada

[4] Univ British Columbia, 3020-950 West 28th Ave, Vancouver, BC V5Z 4H4, Canada

来源：

JOURNAL OF CONTROLLED RELEASE | 2022年 / 350卷

关键词：

Lipid nanoparticle; CRISPR; Ribonucleoprotein; Genome editing; Mouse; Eye; Cornea; tdTomato expression; PAX6; Aniridia; SPONTANEOUS PSEUDOPREGNANCY; PAX6; EYE; EXPRESSION; MANIPULATION; BEVACIZUMAB; MUTATIONS; INJECTION; MUSCLE; VIRUS;

D O I：

10.1016/j.jconrel.2022.08.042

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

CRISPR/Cas9-based genome-editing therapies are poised to change the clinical outcome for many diseases with validated therapeutic targets awaiting an appropriate delivery system. Recent advances in lipid nanoparticle (LNP) technology make them an attractive platform for the delivery of various forms of CRISPR/Cas9, including the efficient and transient Cas9/gRNA ribonucleoprotein (RNP) complexes. In this study, we initially tested our novel LNP platform by delivering pre-complexed RNPs and template DNA to cultured mouse cortical neurons, and obtained successful ex vivo genome editing. We then directly injected LNP-packaged RNPs and DNA template into the mouse cornea to evaluate in vivo delivery. For the first time, we demonstrated wide-spread genome editing in the cornea using our LNP-RNPs. The ability of our LNPs to transfect the cornea highlights the potential of our novel delivery platform to be used in CRISPR/Cas9-based genome editing therapies of corneal diseases.

引用

页码：401 / 413

页数：13

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