Applications of CRISPR-Cas9 mediated genome engineering

Abstract

Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9, is further revolutionizing biology and medical studies. The simplicity of the CRISPR-Cas9 system has enabled its widespread applications in generating germline animal models, somatic genome engineering, and functional genomic screening and in treating genetic and infectious diseases. This technology will likely be used in all fields of biomedicine, ranging from basic research to human gene therapy.

Keywords: Animal model; CRISPR-Cas9; Functional genomic screening; Gene therapy; Genome editing.

Figure 1

The CRISPR-Cas system. The CRISPR-associated endonuclease Cas9 could target specific DNA loci and make double-strand breaks under the guidance of the tracrRNAs:crRNAs duplex. The tracrRNA:crRNA duplex directs Cas9 to use two distinct active sites, RuvC and HNH, and cleave the target DNA complementary to the crRNA, which has an adjacent protospacer-adjacent motif (PAM).

Figure 2

Applications of CRISPR-Cas9 mediated genome engineering.