Research and Therapeutic Approaches in Stem Cell Genome Editing by CRISPR Toolkit

Abstract

The most widely used genome editing toolkit is CRISPR (clustered regularly interspaced short palindromic repeats). It provides the possibility of replacing and modifying DNA and RNA nucleotides. Furthermore, with advancements in biological technology, inhibition and activation of the transcription of specific gene(s) has become possible. Bioinformatics tools that target the evolution of CRISPR-associated protein 9 (Cas9) turn this protein into a vehicle that is specific for a DNA or RNA region with single guide RNA (sgRNA). This toolkit could be used by researchers to investigate the function of stem cell gene(s). Here, in this review article, we cover recent developments and applications of this technique in stem cells for research and clinical purposes and discuss different CRISPR/Cas technologies for knock-out, knock-in, activation, or inhibition of gene expression. Additionally, a comparison of several deliveries and off-target detecting strategies is discussed.

Arp2/3 mediated dynamic lamellipodia of the hPSC colony edges promote liposome-based DNA delivery

“…Gene-edited stem cells can be differentiated to the target cell types for understanding disease mechanisms for developing therapy. Conversely, patient derived induced pluripotent stem cells (iPSCs) with gene mutations can be corrected by CRISPR/Cas9 followed by differentiation to the effected cell type and injection for regenerative medicine (Mollashahi et al, 2023). While these strategies have tremendous clinical benefit and are currently under active research, gene-editing in hPSCs is an extremely inefficient process owing to their poor transfection property (Yang et al, 2013).…”

Neurological Disease Modeling Using Pluripotent and Multipotent Stem Cells: A Key Step towards Understanding and Treating Mucopolysaccharidoses

“…In the specific case of the CRISPR/Cas technology, for example, it has become a relevant platform due to its simple configuration, using advanced algorithms and web-based tools, that allow to analyze its efficiency in recognizing the sequence of interest as well as possible off-targets. Therefore, many advances have been made not only in disease modeling but also in its potential application to gene therapy, namely in combination with stem cells [ 150 ]. Recently, Ahumada-Ayala et al [ 151 ] reviewed a number of studies on therapeutic approaches using CRISPR/Cas, namely in hematological, hemato-oncological, cardiovascular, neurological, infectious, and musculoskeletal diseases.…”