Int J Stem Cells.  2024 Feb;17(1):1-14. 10.15283/ijsc23030.

Recent Research Trends in Stem Cells Using CRISPR/Cas-Based Genome Editing Methods

• Yoon DE
• Lee H
• Kim K

• KMID: 2552013
• DOI: http://doi.org/10.15283/ijsc23030

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR) system, a rapidly advancing genome editing technology, allows DNA alterations into the genome of organisms. Gene editing using the CRISPR system enables more precise and diverse editing, such as single nucleotide conversion, precise knock-in of target sequences or genes, chromosomal rearrangement, or gene disruption by simple cutting. Moreover, CRISPR systems comprising transcriptional activators/repressors can be used for epigenetic regulation without DNA damage. Stem cell DNA engineering based on gene editing tools has enormous potential to provide clues regarding the pathogenesis of diseases and to study the mechanisms and treatments of incurable diseases. Here, we review the latest trends in stem cell research using various CRISPR/Cas technologies and discuss their future prospects in treating various diseases.

Keyword

Clustered regularly interspaced short palindromic repeats; Genome editing; Transcriptional regulator; Stem cell

Figure

• 

  Fig. 1 Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) system in stem cells. CRISPR system could be used for various ex vivo or in vivo stem cell research, such as cell lineage or developmental study, generation of disease research, patient-specific pre-drug screening, and gene therapy. iPSCs: induced pluripotent stem cells, ESCs: embryonic stem cells, RNPs: ribonucleoproteins, AAV: adeno-associated virus.

• 

  Fig. 2 Features and brief action mechanisms of diverse clustered regularly interspaced short palindromic repeats (CRISPR) systems. CRISPR system has been applied in various ways depending on the purpose. Cas nuclease makes random mutation at the cleavage site. The base editor can convert C to T or A to G. Prime editor could insert interested sequence through reverse transcriptase, and CRISPR activation/interference (CRISPRa/i) regulates specific gene expression. sgRNA: single guide RNA, ABE: adenine base editor, PBS: primer-binding site, RTT: Rett syndrome, RT: reverse transcriptase, nCas: nickase Cas, dCas: dead Cas, NHEJ: non-homologous end joining, HDR: homology-directed repair, CBE: cytosine base editor, UGI: uracil glycosylase inhibitor.

Reference

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