CRISPR technologies for stem cell engineering and regenerative medicine

Mu-Nung Hsu; Yu-Han Chang; Vu Anh Truong; Po-Liang Lai; Thị Kieu Nuong Nguyen; Yu-Chen Hu

doi:10.1016/j.biotechadv.2019.107447

CRISPR technologies for stem cell engineering and regenerative medicine

Biotechnol Adv. 2019 Dec;37(8):107447. doi: 10.1016/j.biotechadv.2019.107447. Epub 2019 Sep 9.

Authors

Mu-Nung Hsu¹, Yu-Han Chang², Vu Anh Truong¹, Po-Liang Lai³, Thị Kieu Nuong Nguyen¹, Yu-Chen Hu⁴

Affiliations

¹ Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan.
² Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, Linkou 333, Taiwan; College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
³ Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, Linkou 333, Taiwan; Bone and Joint Research Center, Chang Gung Memorial Hospital, Linkou 333, Taiwan.
⁴ Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan; Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University, Hsinchu 300, Taiwan. Electronic address: yuchen@che.nthu.edu.tw.

PMID: 31513841
DOI: 10.1016/j.biotechadv.2019.107447

Abstract

CRISPR/Cas9 system exploits the concerted action of Cas9 nuclease and programmable single guide RNA (sgRNA), and has been widely used for genome editing. The Cas9 nuclease activity can be abolished by mutation to yield the catalytically deactivated Cas9 (dCas9). Coupling with the customizable sgRNA for targeting, dCas9 can be fused with transcription repressors to inhibit specific gene expression (CRISPR interference, CRISPRi) or fused with transcription activators to activate the expression of gene of interest (CRISPR activation, CRISPRa). Here we introduce the principles and recent advances of these CRISPR technologies, their delivery vectors and review their applications in stem cell engineering and regenerative medicine. In particular, we focus on in vitro stem cell fate manipulation and in vivo applications such as prevention of retinal and muscular degeneration, neural regeneration, bone regeneration, cartilage tissue engineering, as well as treatment of diseases in blood, skin and liver. Finally, the challenges to translate CRISPR to regenerative medicine and future perspectives are discussed and proposed.

Keywords: CRISPR; CRISPRa; CRISPRi; Genome editing; Regenerative medicine; Stem cell.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

CRISPR-Cas Systems*
Clustered Regularly Interspaced Short Palindromic Repeats*
Gene Editing
Regenerative Medicine
Stem Cells