Journal
BIOTECHNOLOGY ADVANCES
Volume 37, Issue 8, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.biotechadv.2019.107447
Keywords
CRISPR; CRISPRa; CRISPRi; Stem cell; Regenerative medicine; Genome editing
Categories
Funding
- Ministry of Science and Technology [MOST 105-2923-E-007-002-MY3, 107-2622-8-007-015, 107-2221-E-007-046-MY3, 107-3017-F-007-002, 108-3017-F-007-003]
- CGMH Intramural Project [CMRPG310181, CMRPG3F1113]
- Frontier Research Center on Fundamental and Applied Sciences of Matters from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education, Taiwan [MOE 108QR001I5]
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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.
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