Exploring the potential of genome editing CRISPR-Cas9 technology.
CRISPR-Cas9 is an RNA-mediated adaptive immune system that protects bacteria and archaea from viruses or plasmids. Herein we discuss the recent development of CRISPR-Cas9 into a key technology for genome editing, targeting, and regulation in a wide range of organisms and cell types. It requires a custom designed single guide-RNA (sgRNA), a Cas9 endonuclease, and PAM sequences in the target region. The sgRNA-Cas9 complex binds to its target and creates a double-strand break (DSB) that can be repaired by non-homologous end joining (NHEJ) or by the homology-directed repair (HDR) pathway, modifying or permanently replacing the genomic target sequence. Additionally, we highlight recent advances in the repurposing of CRISPR-Cas9 for repression, activation, and loci imaging. In this review, we underline the current progress and the future potential of the CRISPR-Cas9 system towards biomedical, therapeutic, industrial, and biotechnological applications.
Institute of Systems and Synthetic Biology, Genopole, CNRS, UEVE, Université Paris-Saclay, 5 rue Henri Desbruères, 91030 Évry, France. Electronic address: vijaisingh15@gmail.com.
Vijai
V
Singh
Institute of Systems and Synthetic Biology, Genopole, CNRS, UEVE, Université Paris-Saclay, 5 rue Henri Desbruères, 91030 Évry, France.
Darren
D
Braddick
Chemical and Synthetic Biology Group, School of Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
Pawan Kumar
PK
Dhar
D000818:Animals; D064113:CRISPR-Cas Systems; D002874:Chromosome Mapping; D000072669:Gene Editing / Q000941:ethics / Q000379:methods* / Q000639:trends; D018390:Gene Targeting; D006801:Humans; D014158:Transcription, Genetic
Activation; CRISPR-Cas9; CRISPRi; Gene therapy; Genome editing; Indel; Repression; sgRNA