富含ω-3型多不饱和脂肪酸基因工程猪研究进展
|
摘要
我国是世界第一大生猪养殖国,猪肉在我国肉类食品组成中占有重要地位,是国人获取动物蛋白的重要来源。猪肉中ω‐6多不饱和脂肪酸(polyunsaturated fatty acids,PUFAs)含量高,而ω‐3 PUFAs含量少,不利于人类健康。基因工程技术可以对特定DNA片段敲除、插入和替换,在改良猪生产性状方面具有巨大潜力,采用基因工程培育富含ω‐3 PUFAs猪具有巨大的市场价值。本文阐述了猪肉ω‐3 PUFAs含量少的原因,并就利用基因工程技术培育富含ω‐3 PUFAs猪的研究进展进行了分析,以期为提升猪肉ω‐3 PUFAs含量的相关研究与实践奠定基础。
China is the largest pig farming country in the world. Pork plays an important role in the composition of meat products in China and is an important source of animal protein for Chinene people. The content of ω‐6 polyunsaturated fatty acids(PUFA)in pork is high,while the content of ω‐3 PUFAs is low,which is not conducive to human health.Genetic engineering technology can knock out,insert and replace specific DNA fragments,and has great potential in improving pig production traits. It is of great market value to breed pigs rich in ω ‐3 PUFAs by genetic engineering technique. In this paper,the reasons for the low content of ω‐3 PUFAs in pork are described,and the research progress of using genetic engineering technique to creat pigs rich in ω‐3 PUFAs is reviewed. This paper lays a foundation for the research and practice of improving the content of pork rich in ω‐3 PUFAs.
China is the largest pig farming country in the world. Pork plays an important role in the composition of meat products in China and is an important source of animal protein for Chinene people. The content of ω‐6 polyunsaturated fatty acids(PUFA)in pork is high,while the content of ω‐3 PUFAs is low,which is not conducive to human health.Genetic engineering technology can knock out,insert and replace specific DNA fragments,and has great potential in improving pig production traits. It is of great market value to breed pigs rich in ω ‐3 PUFAs by genetic engineering technique. In this paper,the reasons for the low content of ω‐3 PUFAs in pork are described,and the research progress of using genetic engineering technique to creat pigs rich in ω‐3 PUFAs is reviewed. This paper lays a foundation for the research and practice of improving the content of pork rich in ω‐3 PUFAs.
引文
[1]Kaur N,Chugh V,Gupta A K.Essential fatty acids as functional components of foodsa review[J].J Food Sci Technol,2014,51(10):22892303.
[2]O'Connell T D,Block R C,Huang S P,et al.ω3Polyunsaturated fatty acids for heart failure:effects of dose on efficacy and novel signaling through free fatty acid receptor 4[J].Journal of Molecular and Cellular Cardiology,2017,(103):7492.
[3]Yu Z,Huang T,Zheng Y,et al.PCSK9 variant,longchain n3 PUFAs,and risk of nonfatal myocardial infarction in Costa Rican Hispanics[J].Am J Clin Nutr,2017,105(5):11981203.
[4]Gu Z N,Wu J S,Wang S H,et al.Polyunsaturated fatty acids affect the localization and signaling of PIP3/AKT in prostate cancer cells[J].Carcinogenesis,2013,34(9):19681975.
[5]Lee S,Lee J,Choi I J,et al.Dietary n3 and n6 polyunsaturated fatty acids,the FADS gene,and the risk of gastric cancer in a Korean population[J].Sci Rep,2018,8(1):3823.
[6]Schwager J,Richard N,Riegger C,et al.ω3 PUFAs and resveratrol differently modulate acute and chronic inflammatory processes[J].BioMed Research International,2015,(2015):111.
[7]Simopoulos A P,Cleland L G,Christiansen E N.Omega6/omega3 essential fatty acid ratio:the scientific evidence[J].Scandinavian Journal of Nutrition,2004,48(1):4950.
[8]Wang K Q,Zhu D,Pang X M,et al.Research on traceability identification for organic pork based on fatty acid differences[J].Guangdong Agricultural Sciences,2014,41(21):106110.王凯强,朱丹,逄秀梅,等.基于脂肪酸差异的有机猪肉溯源识别研究[J].广东农业科学,2014,41(21):106110.
[9]Watts J L.Using Caenorhabditis elegans to uncover conserved functions of omega3 and omega6 fatty acids[J].J Clin Med,2016,5(2):E19.
[10]Spychalla J P,Kinney A J,Browse J.Identification of an animal3 fatty acid desaturase by heterologous expression in Arabidopsis[J].Proceedings of the National Academy of Sciences,1997,94(4):11421147.
[11]Kang Z B,Ge Y,Chen Z,et al.Adenoviral gene transfer of Caenorhabditis elegans n3 fatty acid desaturase optimizes fatty acid composition in mammalian cells[J].Proc Natl Acad Sci USA,2001,98(7):40504054.
[12]Ge Y L,Chen Z H,Kang Z B,et al.Effects of adenoviral gene transfer of C.elegans n3 fatty acid desaturase on the lipid profile and growth of human breast cancer cells[J].Anticancer Res,2002,22(2A):537543.
[13]Liu X F,Wei Z Y,Bai C L,et al.Insights into the function of n3 PUFAs in fat-1 transgenic cattle[J].JLipid Res,2017,58(8):15241535.
[14]Jaenisch R,Mintz B.Simian virus 40 DNA sequences in DNA of healthy adult mice derived from preimplantation blastocysts injected with viral DNA[J].Proc Natl Acad Sci USA,1974,71(4):12501254.
[15]Zhu X X,Quan S N,Huang Y,et al.The application of somatic cell nuclear transfer technology on porcine genetic modification[J].Genomics and Applied Biology,2013,32(6):692699.朱向星,全守能,黄勇,等.体细胞核移植技术在猪基因修饰中的应用[J].基因组学与应用生物学,2013,32(6):692699.
[16]Hao S L,Yan A F,Jiang H,et al.Spermmediated antifreeze protein gene transfer in tiapia[J].Transgenic Res,2014,23(5):857.
[17]Gordon K,Ruddle F H,Gene transfer into mouse embryos[M].Manipulation of Mammalian Development.Springer US,1986,4:136.
[18]Mcevoy T G,Sreenan J M.The efficiency of production,centrifugation,microinjection and transfer of oneand twocell bovine ova in a gene transfer program[J].Theriogenology,1990,33(4):819828.
[19]Hammer R E,Pursel V G,Rexroad C E,et al.Production of transgenic rabbits,sheep and pigs by microinjection[J].Nature,1985,315(6021):680683.
[20]Lavitrano M,Bacci M L,Forni M,et al.Efficient production by spermmediated gene transfer of human decay accelerating factor(hDAF)transgenic pigs for xenotransplantation[J].Proc Natl Acad Sci USA,2002,99(22):1423014235.
[21]Jakobsen J E,Li J,Kragh P M,et al.Pig transgenesis by sleeping beauty DNA transposition[J].Transgenic Res,2011,20(3):533545..
[22]Schnieke A E,Kind A J,Ritchie W A,et al.Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts[J].Science,1997,278(5346):21302133.
[23]Lai L X,Park K W,Cheong H T,et al.Transgenic pig expressing the enhanced green fluorescent protein produced by nuclear transfer using colchicinetreated fibroblasts as donor cells[J].Mol Reprod Dev,2002,62(3):300306.
[24]Tang D S,Jiang H,Liu F,et al.Multilocus,high efficiency gene targeting mediated by zinc finger nucleases[J].Chinese Science Bulletin,2012,57(9):711719.唐冬生,蒋泓,刘芳,等.锌指核酸酶介导的高效多位点基因打靶[J].科学通报,2012,57(9):711719.
[25]Luo T T,Yan A F,Liu L,et al.In vitro study of joint intervention of Ecad and BmiⅠmediated by transcription activatorlike effector nuclease in nasopharyngeal carcinoma[J].Journal of Central South University(Medical Science),2018,43(3):229239.罗婷婷,严爱芬,刘连,等.类转录激活因子效应物核酸酶介导的Ecad和BmiⅠ基因联合干预鼻咽癌的体外研究[J].中南大学学报(医学版),2018,43(3):229239.
[26]Zhu X X,Zhong Y Z,Ge Y W,et al.CRISPR/Cas9mediated generation of Guangxi Bama minipigs harboring three mutations inαsynuclein causing Parkinson's disease[J].Sci Rep,2018,8(1):12420.
[27]Maeder M L,ThibodeauBeganny S,Osiak A,et al.Rapid"opensource"engineering of customized zincfinger nucleases for highly efficient gene modification[J].Mol Cell,2008,31(2):294301.
[28]Sugisaki H,Kanazawa S.New restriction endonucleases from Flavobacterium okeanokoites(Fok I)and Micrococcus luteus(Mlu I)[J].Gene,1981,16(1/2/3):7378.
[29]Zhang Y,Jiang H,Liu F,et al.Electronic design and simulation of specific zinc finger protein[C].IEEEPress,2011,216220.
[30]Liu W X,Yan A F,Liu F,et al.Artificial construction and identification of transcription activatorlike effector nucleases and multiloci gene targeting vector[J].Chinese Journal of Biomedical Engineering,2015,21(5):400406.刘婉霞,严爱芬,刘芳,等.人工转录激活子样效应因子核酸酶与多位点基因打靶载体的构建与鉴定[J].中华生物医学工程杂志,2015,21(5):400406.
[31]Hsu P D,Lander E S,Zhang F.Development and applications of CRISPRCas9 for genome engineering[J].Cell,2014,157(6):12621278.
[32]Horvath P,Barrangou R.CRISPR/Cas,the immune system of bacteria and archaea[J].Science,2010,327(5962):167170.
[33]Wang H Y,Yang H,Shivalila C S,et al.Onestep generation of mice carrying mutations in multiple genes by CRISPR/Casmediated genome engineering[J].Cell,2013,153(4):910918.
[34]Jiang W Y,Bikard D,Cox D,et al.RNAguided editing of bacterial genomes using CRISPRCas systems[J].Nat Biotechnol,2013,31(3):233239.
[35]Platt R J,Chen S D,Zhou Y,et al.CRISPRCas9knockin mice for genome editing and cancer modeling[J].Cell,2014,159(2):440455.
[36]Roper J,Tammela T,Akkad A,et al.Colonoscopybased colorectal cancer modeling in mice with CRISPRCas9 genome editing and organoid transplantation[J].Nat Protoc,2018,13(2):217234.
[37]Ikeda M,Matsuyama S,Akagi S,et al.Publisher correction:correction of a disease mutation using CRISPR/Cas9assisted genome editing in Japanese black cattle[J].Sci Rep,2018,8(1):1470.
[38]Hai T,Teng F,Guo R F,et al.Onestep generation of knockout pigs by zygote injection of CRISPR/Cas system[J].Cell Res,2014,24(3):372375.
[39]Wang K K,Ouyang H S,Xie Z C,et al.Efficient generation of myostatin mutations in pigs using the CRISPR/Cas9 system[J].Sci Rep,2015,(5):16623.
[40]Bi Y Z,Hua Z D,Liu X M,et al.Isozygous and selectable markerfree MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP[J].Sci Rep,2016,(6):31729.
[41]Zheng Q T,Lin J,Huang J J,et al.Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity[J].Proc Natl Acad Sci USA,2017,114(45):E9474E9482.
[42]Wu X,Ouyang H S,Duan B,et al.Production of cloned transgenic cow expressing omega3 fatty acids[J].Transgenic Res,2012,21(3):537543.
[43]Zhang P,Liu P,Dou H W,et al.Handmade cloned transgenic sheep rich in omega3 fatty acids[J].PLoSOne,2013,8(2):e55941.
[44]Lai L X,Kang J X,Li R F,et al.Generation of cloned transgenic pigs rich in omega3 fatty acids[J].Nat Biotechnol,2006,24(4):435436.
[45]Zhang P,Zhang Y D,Dou H W,et al.Handmade cloned transgenic piglets expressing the nematode fat-1gene[J].Cell Reprogram,2012,14(3):258266.
[46]Zhou Y R,Lin Y L,Wu X J,et al.The highlevel accumulation of n3 polyunsaturated fatty acids in transgenic pigs harboring the n3 fatty acid desaturase gene from Caenorhabditis briggsae[J].Transgenic Res,2014,23(1):8997.
[47]Li M J,Ouyang H S,Yuan H M,et al.Sitespecific fat-1 knockin enables significant decrease of n6PUFAs/n3PUFAs ratio in pigs[J].G3(Bethesda),2018,8(5):17471754.
[2]O'Connell T D,Block R C,Huang S P,et al.ω3Polyunsaturated fatty acids for heart failure:effects of dose on efficacy and novel signaling through free fatty acid receptor 4[J].Journal of Molecular and Cellular Cardiology,2017,(103):7492.
[3]Yu Z,Huang T,Zheng Y,et al.PCSK9 variant,longchain n3 PUFAs,and risk of nonfatal myocardial infarction in Costa Rican Hispanics[J].Am J Clin Nutr,2017,105(5):11981203.
[4]Gu Z N,Wu J S,Wang S H,et al.Polyunsaturated fatty acids affect the localization and signaling of PIP3/AKT in prostate cancer cells[J].Carcinogenesis,2013,34(9):19681975.
[5]Lee S,Lee J,Choi I J,et al.Dietary n3 and n6 polyunsaturated fatty acids,the FADS gene,and the risk of gastric cancer in a Korean population[J].Sci Rep,2018,8(1):3823.
[6]Schwager J,Richard N,Riegger C,et al.ω3 PUFAs and resveratrol differently modulate acute and chronic inflammatory processes[J].BioMed Research International,2015,(2015):111.
[7]Simopoulos A P,Cleland L G,Christiansen E N.Omega6/omega3 essential fatty acid ratio:the scientific evidence[J].Scandinavian Journal of Nutrition,2004,48(1):4950.
[8]Wang K Q,Zhu D,Pang X M,et al.Research on traceability identification for organic pork based on fatty acid differences[J].Guangdong Agricultural Sciences,2014,41(21):106110.王凯强,朱丹,逄秀梅,等.基于脂肪酸差异的有机猪肉溯源识别研究[J].广东农业科学,2014,41(21):106110.
[9]Watts J L.Using Caenorhabditis elegans to uncover conserved functions of omega3 and omega6 fatty acids[J].J Clin Med,2016,5(2):E19.
[10]Spychalla J P,Kinney A J,Browse J.Identification of an animal3 fatty acid desaturase by heterologous expression in Arabidopsis[J].Proceedings of the National Academy of Sciences,1997,94(4):11421147.
[11]Kang Z B,Ge Y,Chen Z,et al.Adenoviral gene transfer of Caenorhabditis elegans n3 fatty acid desaturase optimizes fatty acid composition in mammalian cells[J].Proc Natl Acad Sci USA,2001,98(7):40504054.
[12]Ge Y L,Chen Z H,Kang Z B,et al.Effects of adenoviral gene transfer of C.elegans n3 fatty acid desaturase on the lipid profile and growth of human breast cancer cells[J].Anticancer Res,2002,22(2A):537543.
[13]Liu X F,Wei Z Y,Bai C L,et al.Insights into the function of n3 PUFAs in fat-1 transgenic cattle[J].JLipid Res,2017,58(8):15241535.
[14]Jaenisch R,Mintz B.Simian virus 40 DNA sequences in DNA of healthy adult mice derived from preimplantation blastocysts injected with viral DNA[J].Proc Natl Acad Sci USA,1974,71(4):12501254.
[15]Zhu X X,Quan S N,Huang Y,et al.The application of somatic cell nuclear transfer technology on porcine genetic modification[J].Genomics and Applied Biology,2013,32(6):692699.朱向星,全守能,黄勇,等.体细胞核移植技术在猪基因修饰中的应用[J].基因组学与应用生物学,2013,32(6):692699.
[16]Hao S L,Yan A F,Jiang H,et al.Spermmediated antifreeze protein gene transfer in tiapia[J].Transgenic Res,2014,23(5):857.
[17]Gordon K,Ruddle F H,Gene transfer into mouse embryos[M].Manipulation of Mammalian Development.Springer US,1986,4:136.
[18]Mcevoy T G,Sreenan J M.The efficiency of production,centrifugation,microinjection and transfer of oneand twocell bovine ova in a gene transfer program[J].Theriogenology,1990,33(4):819828.
[19]Hammer R E,Pursel V G,Rexroad C E,et al.Production of transgenic rabbits,sheep and pigs by microinjection[J].Nature,1985,315(6021):680683.
[20]Lavitrano M,Bacci M L,Forni M,et al.Efficient production by spermmediated gene transfer of human decay accelerating factor(hDAF)transgenic pigs for xenotransplantation[J].Proc Natl Acad Sci USA,2002,99(22):1423014235.
[21]Jakobsen J E,Li J,Kragh P M,et al.Pig transgenesis by sleeping beauty DNA transposition[J].Transgenic Res,2011,20(3):533545..
[22]Schnieke A E,Kind A J,Ritchie W A,et al.Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts[J].Science,1997,278(5346):21302133.
[23]Lai L X,Park K W,Cheong H T,et al.Transgenic pig expressing the enhanced green fluorescent protein produced by nuclear transfer using colchicinetreated fibroblasts as donor cells[J].Mol Reprod Dev,2002,62(3):300306.
[24]Tang D S,Jiang H,Liu F,et al.Multilocus,high efficiency gene targeting mediated by zinc finger nucleases[J].Chinese Science Bulletin,2012,57(9):711719.唐冬生,蒋泓,刘芳,等.锌指核酸酶介导的高效多位点基因打靶[J].科学通报,2012,57(9):711719.
[25]Luo T T,Yan A F,Liu L,et al.In vitro study of joint intervention of Ecad and BmiⅠmediated by transcription activatorlike effector nuclease in nasopharyngeal carcinoma[J].Journal of Central South University(Medical Science),2018,43(3):229239.罗婷婷,严爱芬,刘连,等.类转录激活因子效应物核酸酶介导的Ecad和BmiⅠ基因联合干预鼻咽癌的体外研究[J].中南大学学报(医学版),2018,43(3):229239.
[26]Zhu X X,Zhong Y Z,Ge Y W,et al.CRISPR/Cas9mediated generation of Guangxi Bama minipigs harboring three mutations inαsynuclein causing Parkinson's disease[J].Sci Rep,2018,8(1):12420.
[27]Maeder M L,ThibodeauBeganny S,Osiak A,et al.Rapid"opensource"engineering of customized zincfinger nucleases for highly efficient gene modification[J].Mol Cell,2008,31(2):294301.
[28]Sugisaki H,Kanazawa S.New restriction endonucleases from Flavobacterium okeanokoites(Fok I)and Micrococcus luteus(Mlu I)[J].Gene,1981,16(1/2/3):7378.
[29]Zhang Y,Jiang H,Liu F,et al.Electronic design and simulation of specific zinc finger protein[C].IEEEPress,2011,216220.
[30]Liu W X,Yan A F,Liu F,et al.Artificial construction and identification of transcription activatorlike effector nucleases and multiloci gene targeting vector[J].Chinese Journal of Biomedical Engineering,2015,21(5):400406.刘婉霞,严爱芬,刘芳,等.人工转录激活子样效应因子核酸酶与多位点基因打靶载体的构建与鉴定[J].中华生物医学工程杂志,2015,21(5):400406.
[31]Hsu P D,Lander E S,Zhang F.Development and applications of CRISPRCas9 for genome engineering[J].Cell,2014,157(6):12621278.
[32]Horvath P,Barrangou R.CRISPR/Cas,the immune system of bacteria and archaea[J].Science,2010,327(5962):167170.
[33]Wang H Y,Yang H,Shivalila C S,et al.Onestep generation of mice carrying mutations in multiple genes by CRISPR/Casmediated genome engineering[J].Cell,2013,153(4):910918.
[34]Jiang W Y,Bikard D,Cox D,et al.RNAguided editing of bacterial genomes using CRISPRCas systems[J].Nat Biotechnol,2013,31(3):233239.
[35]Platt R J,Chen S D,Zhou Y,et al.CRISPRCas9knockin mice for genome editing and cancer modeling[J].Cell,2014,159(2):440455.
[36]Roper J,Tammela T,Akkad A,et al.Colonoscopybased colorectal cancer modeling in mice with CRISPRCas9 genome editing and organoid transplantation[J].Nat Protoc,2018,13(2):217234.
[37]Ikeda M,Matsuyama S,Akagi S,et al.Publisher correction:correction of a disease mutation using CRISPR/Cas9assisted genome editing in Japanese black cattle[J].Sci Rep,2018,8(1):1470.
[38]Hai T,Teng F,Guo R F,et al.Onestep generation of knockout pigs by zygote injection of CRISPR/Cas system[J].Cell Res,2014,24(3):372375.
[39]Wang K K,Ouyang H S,Xie Z C,et al.Efficient generation of myostatin mutations in pigs using the CRISPR/Cas9 system[J].Sci Rep,2015,(5):16623.
[40]Bi Y Z,Hua Z D,Liu X M,et al.Isozygous and selectable markerfree MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP[J].Sci Rep,2016,(6):31729.
[41]Zheng Q T,Lin J,Huang J J,et al.Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity[J].Proc Natl Acad Sci USA,2017,114(45):E9474E9482.
[42]Wu X,Ouyang H S,Duan B,et al.Production of cloned transgenic cow expressing omega3 fatty acids[J].Transgenic Res,2012,21(3):537543.
[43]Zhang P,Liu P,Dou H W,et al.Handmade cloned transgenic sheep rich in omega3 fatty acids[J].PLoSOne,2013,8(2):e55941.
[44]Lai L X,Kang J X,Li R F,et al.Generation of cloned transgenic pigs rich in omega3 fatty acids[J].Nat Biotechnol,2006,24(4):435436.
[45]Zhang P,Zhang Y D,Dou H W,et al.Handmade cloned transgenic piglets expressing the nematode fat-1gene[J].Cell Reprogram,2012,14(3):258266.
[46]Zhou Y R,Lin Y L,Wu X J,et al.The highlevel accumulation of n3 polyunsaturated fatty acids in transgenic pigs harboring the n3 fatty acid desaturase gene from Caenorhabditis briggsae[J].Transgenic Res,2014,23(1):8997.
[47]Li M J,Ouyang H S,Yuan H M,et al.Sitespecific fat-1 knockin enables significant decrease of n6PUFAs/n3PUFAs ratio in pigs[J].G3(Bethesda),2018,8(5):17471754.