CRISPR-Cas9 Gene Editing Technology and Its Research Progress in Poultry

doi:10.13560/j.cnki.biotech.bull.1985.2023-1135

Abstract

Abstract:

The CRISPR-Cas system, found in bacteria or archaea, serves as an adaptive immune system to counteract viral re-invasion by effectively cleaving nucleic acids. By leveraging the characteristics of Cas9 protein and its variants, researchers have devised a range of gene editing techniques that enable gene operations like gene knock-out or knock-in within cells, thereby facilitating genetic diversity in organisms. Additionally, these tools allow the manipulation of DNA or RNA through epigenetic modification to regulate gene expression, rendering them extensively employed in life science research. Poultry, as a significant agricultural species, plays a crucial role in supplying individuals with protein of superior quality. The utilization of CRISPR-Cas9 technology for gene editing and modification in avian genome is presently confined to the research phase, with remarkable advancements observed solely in chicken and quail. This paper presents an overview of CRISPR-Cas9 gene editing technology, encompassing its components, delivery methods, and optimization strategies, while also exploring its potential applications in poultry production and prevention and control of diseases. These insights lay the groundwork for the future development of CRISPR-Cas9 technology in the context of poultry system.

Key words: poultry, CRISPR-Cas9, breeding, diagnosis

XIAO Yi-meng, YANG Wen, CHENG Yi-yi, LUO Gang. CRISPR-Cas9 Gene Editing Technology and Its Research Progress in Poultry[J]. Biotechnology Bulletin, 2024, 40(5): 38-47.

Figures/Tables 3

Fig. 1 Schematic diagram of the CRISPR-Cas9/dCas9 system cutting double-stranded DNA A: Schematic diagram of CRISPR-Cas9 cutting target DNA. Cas9: CRISPR-associated protein 9; RuvC and HNH: structural domains in the Cas9 protein that cleave nucleic acids; PAM sequence: protospacer adjacent motif; sgRNA: single-guide RNA, composed of crRNA（CRISPR RNA）and tracrRNA（trans-activating crRNA）. B and C: Schematic representation of CRISPR-nCas9 cleaving target DNA. nCas9: Cas9 nickase; HNH(H840A): Cas9 mutant with inactivated HNH domain; RuvC（D10A）: Cas9 mutant with inactivated RuvC domain. D: Schematic diagram of CRISPR-dCas9 cutting target DNA. HNH（H840A）and RuvC（D10A）: Cas9 mutants with inactivated HNH and RuvC structural domains

Fig. 2 Delivery of the CRISPR-Cas9 system in avian species A: Schematic diagram of PGC-mediated CRISPR-Cas9 gene editing. U6: gRNA promoter; gRNA: guide RNA; CMV: The promoter of Cas9-encoded genes; GFP: green fluorescent protein; NLS: nuclear localization sequences; PGC: primordial germ cell. B: Schematic diagram of adenovirus-mediated CRISPR-Cas9 gene editing. C: Schematic representation of STAGE-mediated CRISPR-Cas9. STAGE: sperm transfection-assisted gene editing; sgRNA: single guide RNA; Cas 9 mRNA: messenger RNA of Cas 9-encoding genes

Fig. 3 Diagnostic principles of NASBACC based on CRISPR-Cas9 system RBS: Ribosome binding site; Lac Z: β-galactosidase gene

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