Development and potential applications of CRISPR-Cas9 genome editing technology in sarcoma

Abstract

Sarcomas include some of the most aggressive tumors and typically respond poorly to chemotherapy. In recent years, specific gene fusion/mutations and gene over-expression/activation have been shown to drive sarcoma pathogenesis and development. These emerging genomic alterations may provide targets for novel therapeutic strategies and have the potential to transform sarcoma patient care. The RNA-guided nuclease CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein-9 nuclease) is a convenient and versatile platform for site-specific genome editing and epigenome targeted modulation. Given that sarcoma is believed to develop as a result of genetic alterations in mesenchymal progenitor/stem cells, CRISPR-Cas9 genome editing technologies hold extensive application potentials in sarcoma models and therapies. We review the development and mechanisms of the CRISPR-Cas9 system in genome editing and introduce its application in sarcoma research and potential therapy in clinic. Additionally, we propose future directions and discuss the challenges faced with these applications, providing concise and enlightening information for readers interested in this area.

Technological advances in precision medicine and drug development

“…The guide sequences enable fast and cost effective targeting as compared to other specific endonucleases, such as zinc figure nucleases [115]. The use of CRISPR has exploded in the mouse research field where it has been used both in the generation of mouse models of disease and as a treatment of regionalized disease such as the regeneration of muscle function in mdx mice [116,117]. CRISPR recently made headlines when it was first used to introduce alterations in the human embryos in an effort to alter the human β-globin gene, which is mutated in β-thalassemia.…”

CRISPR-Cas9-Mediated Silencing of CD44 in Human Highly Metastatic Osteosarcoma Cells

“…Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9 (CRISPR-Cas9) system has been implemented in a multitude of model organisms and cell types [13-15]. The CRISPR-Cas9 system uses Cas9, which complexes with single guided RNA (sgRNA), to cleave DNA 3-4 base pairs upstream of a protospacer-adjacent motif (PAM) and generate double-strand breaks (DSBs) in a sequence-specific manner [14, 15].…”

“…The CRISPR-Cas9 system uses Cas9, which complexes with single guided RNA (sgRNA), to cleave DNA 3-4 base pairs upstream of a protospacer-adjacent motif (PAM) and generate double-strand breaks (DSBs) in a sequence-specific manner [14, 15]. It is evident that the CRISPR-Cas9 genome editing technology has revolutionized the field of genetic engineering and holds the potential to overcome many of the limitations of earlier techniques to carry out deletions, insertions, translocations, and inversions at specific sites in the DNA of cells [13-15]. …”

Gene drives and the management of agricultural pests

“…CRISPR-Cas9-based gene editing has already provided hope in the treatment of HIV (Kaminski et al 2016), as well as promised to offer a new generation of sickle-cell anemia, hemophilia and cancer treatments (Carroll 2014;Kannan and Ventura 2015;Liu et al 2016;Tang and Shrager 2016). It has shown potential ways to diminish the human suffering caused by malaria and other vector-borne diseases (Ghorbal et al 2014;de Koning-Ward, Gilson, and Crabb 2015;Hammond et al 2016), as well as impacted animal and plant breeding (Jenko et al 2015;Weeks, Spalding, and Yang 2016).…”