CRISPR-Cpf1 Activation of Endogenous BMP4 Gene for Osteogenic Differentiation of Umbilical-Cord-Derived Mesenchymal Stem Cells
| Autor: | Nathaniel S. Hwang, Chris Hyunchul Jo, Taegeun Bae, Junho K. Hur, Ninj Byambasuren, Jae-Hoon Choi, Dokyoung Kim, Seong-Ho Park |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
CRISPR/Cpf1 lcsh:QH426-470 Cellular differentiation Biology Regenerative medicine Article 03 medical and health sciences 0302 clinical medicine Cpf1 Genetics CRISPR lcsh:QH573-671 Bone regeneration Molecular Biology Cell Engineering Regulation of gene expression CRISPR activation lcsh:Cytology Activator (genetics) Mesenchymal stem cell Cell Differentiation Cell biology lcsh:Genetics CRISRP 030104 developmental biology Mesenchymal Stem Cell 030220 oncology & carcinogenesis Molecular Medicine |
| Zdroj: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 309-316 (2020) |
| ISSN: | 2329-0501 |
| Popis: | The CRISPR systems provide powerful genome-editing tools for wide applications in biological and medical research fields. However, the safety issue due to off-target effects of CRISPR has been one of the major hindrances of its application to regenerative medicine. The conventional CRISPR system has the intrinsic danger of inducing unpredictable mutations at non-targeted genomic loci via erroneous double-strand DNA breaks (DSBs). In this study, we demonstrate a safety-enhanced application of a recently discovered CRISPR-Cpf1 for targeted gene activation, without DNA double-strand break, to facilitate osteogenic differentiation of human umbilical-cord-derived mesenchymal stem cells (UC-MSCs). To this end, we developed a catalytically inactive AsCpf1 fused to tripartite transcription activator domain (dAsCpf1-VPR) that can induce upregulation of targeted gene expression in mammalian cells. We observed that the CRISPR-dAsCpf1-VPR activator can be applied to enhance the osteogenic differentiation of human UC-MSCs, via increasing the expression level of endogenous BMP4 gene. The results suggested that the CRISPR-Cpf1 activator provides versatile methods applicable for bone regeneration and regenerative medicine. Graphical Abstract |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|