CRISPR/Cas systems usher in a new era of disease treatment and diagnosis

Abstract

The discovery and development of the CRISPR/Cas system is a milestone in precise medicine. CRISPR/Cas nucleases, base-editing (BE) and prime-editing (PE) are three genome editing technologies derived from CRISPR/Cas. In recent years, CRISPR-based genome editing technologies have created immense therapeutic potential with safe and efficient viral or non-viral delivery systems. Significant progress has been made in applying genome editing strategies to modify T cells and hematopoietic stem cells (HSCs) ex vivo and to treat a wide variety of diseases and disorders in vivo. Nevertheless, the clinical translation of this unique technology still faces many challenges, especially targeting, safety and delivery issues, which require further improvement and optimization. In addition, with the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), CRISPR-based molecular diagnosis has attracted extensive attention. Growing from the specific set of molecular biological discoveries to several active clinical trials, CRISPR/Cas systems offer the opportunity to create a cost-effective, portable and point-of-care diagnosis through nucleic acid screening of diseases. In this review, we describe the development, mechanisms and delivery systems of CRISPR-based genome editing and focus on clinical and preclinical studies of therapeutic CRISPR genome editing in disease treatment as well as its application prospects in therapeutics and molecular detection.

High-Efficiency CRISPR/Cas9-Mediated Correction of a Homozygous Mutation in Achromatopsia-Patient-Derived iPSCs

“…On the other hand, CRISPR/Cas uses a guide RNA (sgRNA) complementary to the target DNA that must, necessarily, be next to a protospacer adjacent motif (PAM) required by Cas endonuclease to cut the DNA. After DNA double-strand breaks (DSBs), cellular machinery primarily repairs DNA via either non-homologous end-joining (NHEJ) or homology-directed repair (HDR) in cases where a donor repair template is provided [ 16 , 24 ]. Notably, HDR is less efficient and is not commonly employed by dividing cells if compared with random repair processes, such as NHEJ or microhomology-mediated end joining (MMEJ) in mammalian cells [ 25 , 26 , 27 , 28 , 29 , 30 ].…”

“…Gene and cell therapy represent promising strategies for next-generation medicine, with a special focus on pathologies caused by genetic defects [ 24 ]. In addition, genome editing constitutes a valuable tool for therapeutic applications, demonstrating striking progress in precision and efficiency [ 24 ].…”

“…Gene and cell therapy represent promising strategies for next-generation medicine, with a special focus on pathologies caused by genetic defects [ 24 ]. In addition, genome editing constitutes a valuable tool for therapeutic applications, demonstrating striking progress in precision and efficiency [ 24 ]. However, despite the notable improvements achieved in recent years, gene therapy is not applicable to all diseases [ 24 ], whereas gene editing tools provide a huge therapeutic potential with the capability of virtually modifying any DNA sequence throughout the genome.…”

“…In addition, genome editing constitutes a valuable tool for therapeutic applications, demonstrating striking progress in precision and efficiency [ 24 ]. However, despite the notable improvements achieved in recent years, gene therapy is not applicable to all diseases [ 24 ], whereas gene editing tools provide a huge therapeutic potential with the capability of virtually modifying any DNA sequence throughout the genome. Thus, gene editing has arisen as a viable approximation for genetic-related diseases because mutated DNA is corrected in situ.…”

“…Thus, gene editing has arisen as a viable approximation for genetic-related diseases because mutated DNA is corrected in situ. In the last years, in vivo CRISPR-based therapeutic strategies have been continuously growing [ 24 , 40 , 41 ]. The majority of human genetic diseases are caused by point mutations [ 25 ]; therefore, gene editing constitutes a promising and important tool for future precise and personalized medicine, and, importantly, for IRDs treatment.…”