猪规模化转基因技术体系构建及其应用

doi:10.3864/j.issn.0578-1752.2014.21.007

Abstract

Abstract: Swine is an important economic animal and pig breeding plays an important role in the development of agriculture in China. At the same time, as a kind of commonly used experimental animals, pigs are similar in anatomy, physiology and genetics to humans and important for life science research. To meet the specific needs of human beings, animal genomes can be modified by transgenic technology using genetic engineering or other experimental techniques, and the target gene can be passed on to the offspring steadily. Therefore, establishment of a scale swine transgenic technology system is essential for animal breeding, gene function and human disease model research. This paper reviewed the research progress of transgenic swine technology system both in China and foreign countries, summarized the principle of the selection of target genes, described the relative merits and applicable scope of specific gene transfer method such as pronuclear microinjection, somatic cell nuclear transfer, zinc-?nger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), clustered regularly interspaced short palindromic repeat/Cas9 (CRISPR/ Cas9), sperm vector and lentiviral vector and so on, expounded the relevant regulatory element and system of gene expression regulation strategy. In recent years, China has made great progress in scale transgenic technology system research since the commencement of National Transgenic Major Projects: including explored a large number of functional genes; built ZFN, TALEN, CRISPR/Cas9 mediated genome modification technology platform; optimized the isolation, culture and screening process of donor cells; improved the somatic cell nuclear transfer and pronuclear microinjection system, and combined them with the novel genome editing techniques for site-specific, safe and efficient integration of exogenous gene; through a further cloning, embryo transfer, artificial insemination and other conventional breeding techniques integrated innovation, finally established the transgenic pig propagation technology system, so that the number of transgenic pigs can be expanded in a short time; provided technical service and trained technical staff for many domestic institutions, made transgenic technology to get a wider range of applications. But a series of problems still exist in the scale swine transgenic technology system of China: the lack of domain genes controlling the transgenic pigs related traits and with independent intellectual property; the transgenic technology supporting system is still not perfect; the potential risks of marker genes; the low integration efficiency and instability of exogenous gene; the inefficiency of gene targeting technology; the low piglets survival rate and so on. All of these are considered to be the bottlenecks of development of scale swine transgenic technology. Therefore, in order to realize the technological breakthrough of scale swine transgenic system, the important functional genes and regulatory element which have independent intellectual property must be explored by related omics tools, construct more efficient, safe and multi-gene transgenic vectors to achieve coordinated and efficient expression of exogenous genes, and improve the related technology in the preparation process of transgenic pigs. In conclusion, the transgenic swine breeding of China will enter a new high-speed development period under the support of scale swine transgenic technology system.

Key words: pig, transgenic system, scale

MU Yu-lian, RUAN Jin-xue, WU Tian-wen, CHENG Ying, WEI Jing-liang, FAN Jun-hua, LI Kui. Construction and Application of a Scale Transgenic Technology System for Pigs[J].Scientia Agricultura Sinica, 2014, 47(21): 4211-4223.

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