CRISPR-Cas9: a promising tool for gene editing on induced pluripotent stem cells

Abstract

Recent advances in genome editing with programmable nucleases have opened up new avenues for multiple applications, from basic research to clinical therapy. The ease of use of the technology—and particularly clustered regularly interspaced short palindromic repeats (CRISPR)—will allow us to improve our understanding of genomic variation in disease processes via cellular and animal models. Here, we highlight the progress made in correcting gene mutations in monogenic hereditary disorders and discuss various CRISPR-associated applications, such as cancer research, synthetic biology, and gene therapy using induced pluripotent stem cells. The challenges, ethical issues, and future prospects of CRISPR-based systems for human research are also discussed.

CRISPR-Cas9 targeting of MMP13 in human chondrocytes leads to significantly reduced levels of the metalloproteinase and enhanced type II collagen accumulation

“…It induces double strand breaks (DSBs) which repair through non-homologous end-joining (NHEJ), or in the presence of a suitable DNA template, by homology-directed repair (HDR). The CRISPR-Cas9 system thus enables highly specific gene editing giving it great potential for correction of genetic (particularly monogenic) diseases 4,5 , or for revolutionising cell-based therapies. Genome editing of the germline in humans is obviously fraught with ethical considerations.…”

Risk associated with off-target plant genome editing and methods for its limitation

“…In the human germline engineering field, the use of the CRISPR/Cas9 system faces clear ethical issues because of unanticipated effects of editing [100], and this concern will bleed over to considerations of crop genome editing. Clearly, more research and continued public discourse and deliberation about gene-editing technology and concerns from various perspectives will be needed for both human therapeutic use and crop improvement.…”

Next Steps in Cancer Therapy: Integration of Gene Editing in Inducible Pluripotent Stem Cells

“…Interestingly, while several groups are comparing TALEN and CRISPR independently, others are combining them to develop inducible-CRISPR methods, which can promote genetic modification at multiple loci following iPSC stimulation. The application of these technologies is crucial for mechanistic interrogation of complex and pleiotropic genetic mutations that are often observed in cancer genetics (8,9). Additionally, efforts have been undertaken to improve CRISPR/Cas9 technology for use in the clinical field to address the off-target concerns and to investigate the integration of the entire Cas9 plasmid construct into the targeted genomic loci (10).…”