Efficient in Vivo Gene Editing Using Ribonucleoproteins in Skin Stem Cells of Recessive Dystrophic Epidermolysis Bullosa Mouse Model

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2017 Feb 1

PMID 28137859

Citations 50

Authors

Wenbo Wu

Zhiwei Lu

Fei Li

Wenjie Wang

Nannan Qian

Jinzhi Duan

Yu Zhang

Fengchao Wang

Ting Chen

Affiliations

Soon will be listed here.

Abstract

The prokaryotic CRISPR/Cas9 system has recently emerged as a powerful tool for genome editing in mammalian cells with the potential to bring curative therapies to patients with genetic diseases. However, efficient in vivo delivery of this genome editing machinery and indeed the very feasibility of using these techniques in vivo remain challenging for most tissue types. Here, we show that nonreplicable Cas9/sgRNA ribonucleoproteins can be used to correct genetic defects in skin stem cells of postnatal recessive dystrophic epidermolysis bullosa (RDEB) mice. We developed a method to locally deliver Cas9/sgRNA ribonucleoproteins into the skin of postnatal mice. This method results in rapid gene editing in epidermal stem cells. Using this method, we show that Cas9/sgRNA ribonucleoproteins efficiently excise exon80, which covers the point mutation in our RDEB mouse model, and thus restores the correct localization of the collagen VII protein in vivo. The skin blistering phenotype is also significantly ameliorated after treatment. This study provides an in vivo gene correction strategy using ribonucleoproteins as curative treatment for genetic diseases in skin and potentially in other somatic tissues.

Citing Articles

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Current Status of Biomedical Products for Gene and Cell Therapy of Recessive Dystrophic Epidermolysis Bullosa.

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