用于器官移植的基因修饰猪的研究进展

张军方; 徐佳; 蔡志明; 戴一凡; 牟丽莎

doi:10.3969/j.issn.1674-7445.2017.02.014

用于器官移植的基因修饰猪的研究进展

doi: 10.3969/j.issn.1674-7445.2017.02.014

510275 广州，中山大学（张军方、牟丽莎）；深圳市第二人民医院（张军方、徐佳、蔡志明、牟丽莎）；南京医科大学（戴一凡）

基金项目:

国家自然科学基金项目青年基金 81502685

广东省自然科学基金项目 2016A030310179

深圳市高水平医学学科建设专项基金 2016031638

深圳市三名工程；深圳市科创委学科布局项目 JCYJ20160229204849975

深圳市科创委企业工程中心项目 GCZX2015043017281705

详细信息

通讯作者:
戴一凡, Email: daiyifan@njmu.edu.cn

牟丽莎, Email: molly__molly@163.com

中图分类号: R617, R-332, Q78
计量
- 文章访问数: 101
- HTML全文浏览量: 48
- PDF下载量: 9
- 被引次数: 0
出版历程
- 收稿日期: 2017-01-03
- 网络出版日期: 2021-01-19
- 刊出日期: 2017-03-15

摘要

摘要: 器官的严重短缺是开展移植手术面临的主要难题。经过基因修饰改造的低免疫源性猪有望成为合适的异种器官移植供体。人工核酸内切酶介导的基因组编辑技术，包括ZFNs、TALENs和CRISPR/Cas9技术，极大地促进了异种器官移植的发展，并且可用于高效定向的遗传改造。目前，基因修饰猪的研究正在朝临床应用的方向飞速发展。本文综述了利用基因组编辑技术培育的基因修饰猪的研究进展。
- 异种移植 /
- 锌指结构核酸酶 /
- 核酸内切酶 /
- 脱靶效应

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表 1 传统基因编辑技术在转基因修饰猪中的应用

Table 1. Application of traditional gene editing technology in transgenic pigs

研究作者	发表年份	靶基因	技术方案	基因编辑技术
Fodor WL, et al^[5]	1994	Human CD59 ⁺	H2kb-hCD59 DNA for embryo injection	Random integration
Cozzi E, et al^[6]	1995	Human CD55 ⁺	6.5 kilobase DNA construct with the 4 kb hDAF	Random integration
Osman N, et al^[7]	1997	Human GLA ⁺	pH2kb-hHT	Random integration
Costa C, et al^[8]	1999	Human H-transferase ⁺	H2Kb-HT construct	Random integration
Miyagawa S, et al^[9]	2001	Human CD46⁺	60 kb genomic construct with CD46	Random integration
Miyagawa S, et al^[9]	2001	Human GnT-Ⅲ⁺	pCX vector-GnT-Ⅲ	Random integration
Lai L, et al^[10]	2002	GGTA1 ^－/+	—	Random integration
Phelps CJ, et al^[11]	2003	GGTA1 ^－/－	back cross	Random integration
Klose R, et al^[12]	2005	Human TRAIL ⁺	TRAIL in pUCH2XXS-TRAIL	Random integration
Wu G, et al^[13]	2007	Human DAF and MCP ⁺	—	Random integration
Phelps CJ, et al^[14]	2009	Porcine CTLA4-Ig ⁺	pCTLA4-Ig with CHS and CH3 linker of IgG1	Random integration
Petersen B, et al^[15]	2009	Human thrombomodulin ⁺	hTM in pEGFP-N1	Random integration
Weiss EH, et al^[16]	2009	HLA-E/Human Beta-2-microglobulin ⁺	HLA-B7 in pCR2.1-TOPO	Random integration
Oropeza M, et al^[17]	2009	Human A20 ⁺	pCAGGSEhA20-IRESNEO	Random integration
Yazaki S, et al^[18]	2009	Endo-B-Galactosidase t and Human CD55 ⁺	pCAG-hDAF-p (A) and pCAGGS/GT t EdnoGalC-neo	Random integration
Hara H, et al^[19]	2010	CⅡTA-DN ⁺	—	Random integration
Shudo K, et al^[20]	2010	Human Fas Ligand ⁺	pcDNA/FasL	Random integration
Cho B, et al^[21]	2011	Human TNFRI-Fc ⁺	shTNFRI-Fc	Random integration
Yeom HJ, et al^[22]	2012	Human heme oxygenase 1 ⁺	SV40-hHO-I	Random integration
Wheeler DG, et al^[23]	2012	Human CD39 ⁺	H2Kb-hCD39	Random integration
Klymiuk N, et al^[24]	2012	LEA29Y ⁺	LEA29Y in B-cell specific expression vector	Random integration

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表 2 人工核酸酶技术在转基因修饰猪中的应用

Table 2. Application of artificial nucleic acid enzyme in transgenic modified pig

研究作者	发表年份	靶基因	基因编辑技术
Lutz AJ, et al^[28]	2013	GGTA1^－/－ CMAH ^－/－	ZFN
Li P, et al^[35]	2014	GGTA1^－/－ CMAH ^－/－ iGb3S ^－/－	CRISPR/Cas9
Li P, et al ^[35]	2014	GGTA1^－/－ CMAH ^－/－	CRISPR/Cas9
Li P, et al ^[35]	2014	GGTA1^－/－	CRISPR/Cas9
Li P, et al^[35]	2014	GGTA1^－/－ iGb3S^－/－	CRISPR/Cas9
Reyes LM, et al^[36]	2014	GGTA1^－/－ SLA-1, -2, -3^－/－	CRISPR/Cas9
Reyes LM, et al^[36]	2015	ASGR^－/－	TALEN
Estrada JL, et al^[37]	2015	GGTA1^－/－ CMAH^－/－ B4GalNT2^－/－	CRISPR/Cas9

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表 3 目前用于异种移植的基因修饰猪的遗传改造情况

Table 3. Current situation of genetically modified pigs for xenotransplantation

转基因猪改进策略	修饰的基因及方法
Gal or non-Gal antigen ‘masking’ or deletion	Human H-transferase gene expression (expression of blood type O antigen)
	Endo-β-galactosidase C (reduction of Gal antigen expression)
	α1, 3-galactosyltransferase gene knockout (GTKO)
	Cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene knockout (CMAHKO)
	β1, 4-N-acetylgalactosaminyltransferase (β4GalNT2) gene knockout (β4GalNT2KO)
	CD46 (membrane co-factor protein)
Complement regulation by human complement-regulatory gene expression	CD55 (decay-accelerating factor)
	CD59 (protectin or membrane inhibitor of reactive lysis)
	von Willebrand factor (vWF)-deficient (natural mutant)
Anticoagulation and anti-inflammatory gene expression or deletion	Human tissue factor pathway inhibitor (TFPI)
	Human thrombomodulin
	Human endothelial protein C receptor (EPCR)
	Human ectonucleoside triphosphate diphosphohydrolase-1 (CD39)
	CIITA-DN (MHC class Ⅱ transactivator knockdown, resulting in swine leukocyte antigen class Ⅱ knockdown)
Suppression of cellular immune response by gene expression or downregulation	Class Ⅰ MHC-knockout (MHC-IKO)
	HLA-E/human β2-microglobulin (inhibits human natural killer cell cytotoxicity)
	Human FAS ligand (CD95L)
	Human N-acetylglucosaminyltransferase Ⅲ (GnT-Ⅲ) gene
	Porcine CTLA4-Ig (cytotoxic T lymphocyte antigen 4 or CD152)
	Human TRAIL (tumour necrosis factor-α-related apoptosis-inducing ligand)
	Human tumour necrosis factor-α-induced protein 3 (A20)
Anticoagulation, anti-inflammatory and anti-apoptotic gene expression	Human haem oxygenase-1 (HO-1)
	Human CD47 (species-specific interaction with SIRP-α inhibits phagocytosis)
	Porcine asialoglycoprotein receptor 1 gene-knockout (ASGR1-KO; decreases platelet phagocytosis)
	Human signal regulatory protein-α (SIRPα; decreases platelet phagocytosis by ‘self’ recognition)
	PERV siRNA
Prevention of porcine endogenous retrovirus (PERV) activation	Genome-wide inactivation

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