Smits Jos P H, Meesters Luca D, Maste Berber G W, Zhou Huiqing, Zeeuwen Patrick L J M, van den Bogaard Ellen H
Department of Dermatology, Radboud University Medical Center (Radboudumc), Radboud Institute for Molecular Life Sciences (RIMLS), Nijmegen, The Netherlands.
Department of Molecular Developmental Biology, Faculty of Science, Radboud University, Radboud Institute for Molecular Life Sciences (RIMLS), Nijmegen, The Netherlands.
JID Innov. 2021 Dec 1;2(2):100082. doi: 10.1016/j.xjidi.2021.100082. eCollection 2022 Mar.
CRISPR-Cas9 is the most straightforward genome-editing tool to date. However, its implementation across disciplines is hampered by variable genome-editing efficiencies, reduced cell viability, and low success rates in obtaining clonal cell lines. This review aims to recognize all CRISPR-Cas9‒related work within the experimental dermatology field to identify key factors for successful strategies in the different keratinocyte (KC) cell sources available. On the basis of these findings, we conclude that most groups use immortalized KCs for generating knockout KCs. Our critical considerations for future studies using CRISPR-Cas9, both for fundamental and clinical applications, may guide implementation strategies of CRISPR-Cas9 technologies in the (experimental) dermatology field.
CRISPR-Cas9是迄今为止最直接的基因组编辑工具。然而，其在各学科中的应用受到基因组编辑效率参差不齐、细胞活力降低以及获得克隆细胞系成功率低的阻碍。本综述旨在梳理实验性皮肤病学领域内所有与CRISPR-Cas9相关的研究工作，以确定在不同可用角质形成细胞（KC）细胞来源中成功策略的关键因素。基于这些发现，我们得出结论，大多数研究小组使用永生化KC来生成基因敲除KC。我们对未来CRISPR-Cas9基础和临床应用研究的关键思考，可能会指导CRISPR-Cas9技术在（实验性）皮肤病学领域的实施策略。
