Abstract Precise genome editing technologies not only play important roles in biological research and crop improvement but also are a potential avenue for therapy of genetic diseases. Clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins (CRISPR/Cas) has been developed as a new targeted genome editing tool since the last 2 years. Because it's flexible, efficient, cheap and easy to operate, it quickly surpass the previous technologies to become the hottest genome editing tool. Up to now, CRISPR/Cas has been applied in a wide variety of cells and organisms such as bacteria, mammalian cells, Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster, Brachydanio rerio, Arabidopsis thaliana, Oryza sativa and so on. Furthermore, the CRISPR-Cas9 system has broad application prospects in functional genomic screens, transcriptional modulation, epigenetic control and live imaging of DNA loci. It will undoubtedly transform biological research and spur the development of biotechnology. Here, we review the biological basis of CRISPR/Cas DNA-targeting platform, with an emphasis on means to improve target specificity, provide an update of recent developments and applications, and present the challenges and future directions for this platform to provide some useful references for the later research and application of this field.
