CRISPR/Cas9 in Genome Editing and Beyond

Abstract

The Cas9 protein (CRISPR-associated protein 9), derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is emerging as a powerful tool for engineering the genome in diverse organisms. As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence, and its development as a tool has made sequence-specific gene editing several magnitudes easier. The nuclease-deactivated form of Cas9 further provides a versatile RNA-guided DNA-targeting platform for regulating and imaging the genome, as well as for rewriting the epigenetic status, all in a sequence-specific manner. With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics. In this review, we describe the current models of Cas9 function and the structural and biochemical studies that support it. We focus on the applications of Cas9 for genome editing, regulation, and imaging, discuss other possible applications and some technical considerations, and highlight the many advantages that CRISPR/Cas9 technology offers.

Pathogenic potential and virulence genotypes of intestinal and faecal isolates of porcine post-weaning enteropathogenic Escherichia coli

“…It is hypothesized that the genetic modification negatively influenced the ability of the development of the roots of rice plants, thereby reducing their yield under field conditions (Dubock, 2014). If indeed this is the case, the newly developed genome editing techniques (such as CRISPR) may save the golden rice project, because they target very specific regions in the DNA, in contrast to the 'classical' GMO method, where the point of gene introduction is random (Wang et al, 2016). The CRISPR technique is based on the immune procedure of bacteria or many archaea against viral (phage) infections and relatively simple and inexpensive.…”

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

“…Only recently discovered, CRISPR/Cas9 technology has already been enhanced to the point of fulfilling most of the genome editing and gene regulation currently demanded, ranging from the ability to perform multiple gene insertions, gene knockouts, combinatorial libraries, to advanced fine-tuning of biosynthetic pathways [23,39,47,95]. However, off-targeting remains an important limitation to the Retrotransposons are composed of similar DNA sequences, which ultimately allows to generate a promiscuous gRNA able to target several of these targets at once.…”

Combinations of chromosome transfer and genome editing for the development of cell/animal models of human disease and humanized animal models

“…These genome editing technologies allow KO or knock in cultures cells as well as animals to be produced. Details of the genome editing technologies are described in other reviews [36,37]. Here, we describe the development of previously reported chromosome transfer-related technologies, and a new generation of chromosome transfer technology combined with genome editing techniques.…”