Current updates of CRISPR/Cas9-mediated genome editing and targeting within tumor cells:an innovative strategy of cancer management

Clustered regularly interspaced short palindromic repeats-associated protein(CRISPR/Cas9),an adaptive microbial immune system,has been exploited as a robust,accurate,efficient and programmable method for genome targeting and editing.This innovative and revolutionary technique can play a significant role in animal modeling,in vivo genome therapy,engineered cell therapy,cancer diagnosis and treatment.The CRISPR/Cas9 endonuclease system targets a specific genomic locus by single guide RNA(sgRNA),forming a heteroduplex with target DNA.The Streptococcus pyogenes Cas9/sgRNA:DNA complex reveals a bilobed architecture with target recognition and nuclease lobes.CRISPR/Cas9 assembly can be hijacked,and its nanoformulation can be engineered as a delivery system for different clinical utilizations.However,the efficient and safe delivery of the CRISPR/Cas9 system to target tissues and cancer cells is very challenging,limiting its clinical utilization.Viral delivery strategies of this system may have many advantages,but disadvantages such as immune system stimulation,tumor promotion risk and small insertion size outweigh these advantages.Thus,there is a desperate need to develop an efficient non-viral physical delivery system based on simple nanoformulations.The delivery strategies of CRISPR/Cas9 by a nanoparticle-based system have shown tremendous potential,such as easy and large-scale production,combination therapy,large insertion size and efficient in vivo applications.This review aims to provide in-depth updates on Streptococcus pyogenic CRISPR/Cas9 structure and its mechanistic understanding.In addition,the advances in its nanoformulation-based delivery systems,including lipid-based,polymeric structures and rigid NPs coupled to special ligands such as aptamers,TAT peptides and cell-penetrating peptides,are discussed.Furthermore,the clinical applications in different cancers,clinical trials and future prospects of CRISPR/Cas9 delivery and genome targeting are also discussed.