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猪的基因组编辑

Genome Editing of Pig.

作者信息

Watanabe Masahito, Nagashima Hiroshi

机构信息

Meiji University International Institute for Bio-Resource Research, Kawasaki, 214-8571, Japan.

Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, 214-8571, Japan.

出版信息

Methods Mol Biol. 2017;1630:121-139. doi: 10.1007/978-1-4939-7128-2_11.

Abstract

Pigs are important livestock for food and have been used in various biomedical studies, particularly translational research, as experimental animals because of their anatomical and physiological similarity to humans. The recent development of genome editing techniques, such as ZFN, TALEN, and CRISPR/Cas9, has rapidly expanded the use of genome editing tools in a variety of animals, resulting in the relatively easy and efficient generation of gene knock-out pigs. In the past few years, there has been a sustained increase in reports describing the development of genetically modified pigs. This chapter introduces our workflow for establishing the genetically modified cells (nuclear donor cells) necessary to create gene knock-out pigs using somatic cell nuclear transfer and focuses on the actual generation of gene knock-out pigs using a cytoplasmic injection method.

摘要

猪是重要的食用家畜,由于其在解剖学和生理学上与人类相似,已被用作各种生物医学研究,特别是转化研究中的实验动物。最近,诸如锌指核酸酶(ZFN)、转录激活样效应因子核酸酶(TALEN)和规律成簇间隔短回文重复序列/Cas9(CRISPR/Cas9)等基因组编辑技术的发展,迅速扩大了基因组编辑工具在各种动物中的应用,从而相对容易且高效地培育出基因敲除猪。在过去几年中,描述转基因猪培育的报道持续增加。本章介绍了我们使用体细胞核移植创建基因敲除猪所需的转基因细胞(核供体细胞)的建立流程,并重点介绍了使用细胞质注射法实际培育基因敲除猪的过程。

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