基因敲除技术在大型家畜中的应用

doi:10.11843/j.issn.0366-6964.2014.01.001

摘要/Abstract

摘要：

基因敲除作为一种重要的生物技术，被广泛应用于当今生物学、医学研究中，并逐渐扩展到农业、畜牧业和兽医学等研究领域。与传统的同源重组介导的基因敲除相比，近几年发展起来的几种新技术包括RNA干扰（RNAi）、锌指核酶技术（ZFNs）、转录激活子类似的效应子核酶技术（TALENs）和成簇的规律间隔性短回文重复序列 (CRISPR/Cas）等，能够在复杂的基因组和RNA水平引入精细的改变或基因沉默。这些技术在大型家畜的疾病防治、性状改良等研究中发挥着积极的促进作用。随着这些技术的不断发展，以及新技术的开发和应用，会有更多的基因敲除手段应用到生产实践中，迅速地推动畜牧业的发展。本文主要针对RNA干扰（RNAi），锌指核酶技术（ZFNs），转录激活子类似的效应子核酶技术（TALENs）3种新兴技术在猪、牛、羊等大型家畜中的应用进行简要论述。

Abstract:

As a powerful tool，gene knockout technology has been widely used in biological and medical researches，and gradually utilized to other fields such as agriculture，livestock husbandry and veterinary science.Comparing to traditional homologous recombination，several novel techniques including RNA interference (RNAi)，zinc finger nuclease (ZFNs)，transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR/Cas），which have been developed in recent years，can introduce delicate alteration and silencing in complex genomic DNA and RNA.These techniques are facilitating the disease prevention and trait improvement of big domestic animals.The improvement of these techniques and emergence of new techniques，and their application in domestic animals will push the development of livestock husbandry.This article is a brief overview focusing on the application of new gene knockout techniques include RNAi，ZFNs and TALENs in big domestic animals such as swine，bovine and caprine.

中图分类号:

S814.8" target="_blank"> S814.8

赵宇航，梁浩，刘明秋，李雪玲. 基因敲除技术在大型家畜中的应用[J]. 畜牧兽医学报, 2014, 45(1): 1-8.

ZHAO Yu-hang，LIANG Hao，LIU Ming-qiu，LI Xue-ling. The Application of Gene Knockout Technologies in Big Domestic Animals[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(1): 1-8.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2014.01.001

https://www.xmsyxb.com/CN/Y2014/V45/I1/1

参考文献

［1］SIOMI H，SIOMI M C.On the road to reading the RNA-interference code［J］.Nature，2009，457(7228):396-404.

［2］HIRANO T，YAMAUCHI N，SATO F，et al.Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes［J］. J Reprod Dev，2004，50(5):599-603.

［3］KARLAS A，KURTH R，DENNER J.Inhibition of porcine endogenous retroviruses by RNA interference:increasing the safety of xenotransplantation［J］.Virology，2004，325(1):18-23.

［4］RAMSOONDAR J，VAUGHT T，BALL S，et al.Production of transgenic pigs that express porcine endogenous retrovirus small interfering RNAs［J］.Xenotransplantation，2009，16(3):164-180.

［5］HE Y X，HUA R H，ZHOU Y J，et al.Interference of porcine reproductive and respiratory syndrome virus replication on MARC-145 cells using DNA-based short interfering RNAs［J］.Antiviral Res，2007，74(2):83-91.

［6］LI G，JIANG P，LI Y，et al.Inhibition of porcine reproductive and respiratory syndrome virus replication by adenovirus-mediated RNA interference both in porcine alveolar macrophages and swine［J］.Antiviral Res，2009，82(3):157-165.

［7］BAO Y，GUO Y，ZHANG L，et al.Inhibition of porcine reproductive and respiratory syndrome virus replication by RNA interference in MARC-145 cells［J］.Mol Biol Rep，2012，39(3):2515-2522.

［8］ZHOU F，LIANG S，CHEN A H，et al.A transgenic Marc-145 cell line of piggyBac transposon-derived targeting shRNA interference against porcine reproductive and respiratory syndrome virus［J］. Vet Res Commun，2012，36(2):99-105.

［9］LIU J，CHEN I，CHUA H，et al.Inhibition of porcine circovirus type 2 replication in mice by RNA interference［J］.Virology，2006，347(2):422-433.

［10］SUN M，LIU X，CAO S，et al.Inhibition of porcine circovirus type 1 and type 2 production in PK-15 cells by small interfering RNAs targeting the Rep gene［J］.Vet Microbiol，2007，123(1-3):203-209.

［11］ZHOU J F，HUA X G，CUI L，et al.Effective inhibition of porcine transmissible gastroenteritis virus replication in ST cells by shRNAs targeting RNA-dependent RNA polymerase gene［J］.Antiviral Res，2007，74(1):36-42.

［12］LI J，GUO H，SHI Z，et al.In vitro inhibition of CSFV replication by retroviral vector-mediated RNA interference［J］.J Virol Methods，2010，169(2):316-321.

［13］PORNTRAKULPIPAT S，SUPANKONG S，CHATCHAWANCHONTEERA A，et al.RNA interference targeting nucleocapsid protein (C) inhibits classical swine fever virus replication in SK-6 cells［J］.Vet Microbiol，2010，142(1-2):41-44.

［14］LI J，DAI Y，LIU S，et al.In vitro inhibition of CSFV replication by multiple siRNA expression［J］.Antiviral Res，2011，91(2):209-216.

［15］LV K，GUO Y，ZHANG Y，et al.Transient inhibition of foot-and-mouth disease virus replication by siRNAs silencing VP1 protein coding region［J］.Res Vet Sci，2009，86(3):443-452.

［16］CONG W，JIN H，JIANG C，et al.Attenuated Salmonella choleraesuis-mediated RNAi targeted to conserved regions against foot-and-mouth disease virus in guinea pigs and swine［J］.Vet Res，2010，41(3):30-43.

［17］LUO J，DU J，GAO S，et al.Lentviral-mediated RNAi to inhibit target gene expression of the porcine integrin alphav subunit，the FMDV receptor，and against FMDV infection in PK-15 cells［J］.Virol J，2011，8(1):428-437.

［18］CHEN J W，ZHANG Y，ZHANG Y L，et al.Construction of multiple shRNAs expression vector that inhibits FUT1 gene expression and production of the transgenic SCNT embryos in vitro［J］.Mol Biol Rep，2012，40(3):2243-2252.

［19］STEWART C K，LI J，GOLOVAN S P.Adverse effects induced by short hairpin RNA expression in porcine fetal fibroblasts［J］.Biochem Biophys Res Commun，2008，370(1):113-117.

［20］CHI H，SHINOHARA M，YOKOMINE T，et al.Successful suppression of endogenous alpha-1,3-galactosyltransferase expression by RNA interference in pig embryos generated in vitro［J］.J Reprod Dev，2012，58(1):69-76.

［21］GUO X K，ZHANG Q，GAO L，et al.Increasing expression of microRNA 181 inhibits porcine reproductive and respiratory syndrome virus replication and has implications for controlling virus infection［J］.J Virol，2013，87(2):1159-1171.

［22］XIA B，SONG H，CHEN Y，et al.Efficient inhibition of porcine reproductive and respiratory syndrome virus replication by artificial microRNAs targeting the untranslated regions［J］.Arch Virol，2013，158(1):55-61.

［23］LAMBETH L S，MOORE R J，MURALITHARAN M S，et al.Suppression of bovine viral diarrhea virus replication by small interfering RNA and short hairpin RNA-mediated RNA interference［J］.Vet Microbiol，2007，119(2-4):132-143.

［24］CHUANG S T，JI W T，CHEN Y T，et al.Suppression of bovine ephemeral fever virus by RNA interference［J］.J Virol Methods，2007，145(1):84-87.

［25］GOLDING M C，LONG C R，CARMELL M A，et al.Suppression of prion protein in livestock by RNA interference［J］.Proc Natl Acad Sci U S A，2006，103(14):5285-5290.

［26］SUTOU S，KUNISHI M，KUDO T，et al.Knockdown of the bovine prion gene PRNP by RNA interference (RNAi) technology［J］.BMC Biotechnol，2007，7:44-54.

［27］WONGSRIKEAO P，SUTOU S，KUNISHI M，et al.Combination of the somatic cell nuclear transfer method and RNAi technology for the production of a prion gene-knockdown calf using plasmid vectors harboring the U6 or tRNA promoter［J］.Prion，2011，5(1):39-46.

［28］WANG H，WU J，LIU X，et al.Identification of short hairpin RNA targeting foot-and-mouth disease virus with transgenic bovine fetal epithelium cells［J］.PLoS ONE，2012，7(8):e42356.

［29］WANG H，LIU X，WU J，et al.Bovine fetal epithelium cells expressing shRNA targeting viral VP1 gene resisted against foot-and-mouth disease virus［J］.Virology，2013，439(2):115-121.

［30］TRIPATHI A K，RAMANI U V，PATEL A K，et al.Short hairpin RNA-induced myostatin gene silencing in caprine myoblast cells in vitro［J］.Appl Biochem Biotechnol，2013，169(2):688-694.

［31］HU S，NI W，SAI W，et al.Knockdown of myostatin expression by RNAi enhances muscle growth in transgenic sheep［J］.PLoS ONE，2013，8(3):e58521.

［32］TANG D，ZHU H，WU J，et al.Silencing myostatin gene by RNAi in sheep embryos［J］.J Biotechnol，2012，158(3):69-74.

［33］ZHANG S，XIONG K，XIE Z，et al.Stable silencing of beta-lactoglobulin (BLG) gene by lentivirus-mediated RNAi in goat fetal fibroblasts［J］.Genet Mol Biol，2012，35(3):680-685.

［34］JABED A，WAGNER S，MCCRACKEN J，et al.Targeted microRNA expression in dairy cattle directs production of beta-lactoglobulin-free，high-casein milk［J］.Proc Natl Acad Sci U S A，2012，109(42):16811-16816.

［35］LU C，YANG R，SHEN B，et al.RNA interference-mediated knockdown of DGAT1 decreases triglyceride content of bovine mammary epithelial cell line［J］.Gene Expr，2012，15(5-6):199-206.

［36］GANTIER M P，WILLIAMS B R G.The response of mammalian cells to double-stranded RNA［J］.Cytokine Growth Factor Rev，2007，18(5-6):363-371.

［37］URNOV F D，MILLER J C，LEE Y L，et al.Highly efficient endogenous human gene correction using designed zinc-finger nucleases［J］. Nature，2005，435(7042):646-651.

［38］HAUSCHILD Q J，PETERSEN B，COST G，et al.Gene knockout and knockin by zinc-finger nucleases:current status and perspectives［J］.Cell Mol Life Sci，2012:1-15.

［39］WATANABE M，UMEYAMA K，MATSUNARI H，et al.Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases［J］.Biochem Biophys Res Commun，2010，402(1):14-18.

［40］WHYTE J J，ZHAO J，WELLS K D，et al.Gene targeting with zinc finger nucleases to produce cloned eGFP knockout pigs［J］.Mol Reprod Dev，2011，78(1):2.

［41］YANG D，YANG H，LI W，et al.Generation of PPAR gamma mono-allelic knockout pigs via zinc-finger nucleases and nuclear transfer cloning［J］.Cell Res，2011，21(6):979-982.

［42］HAUSCHILD J，PETERSEN B，SANTIAGO Y，et al.Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases［J］. Proc Natl Acad Sci U S A，2011，108(29):12013-12017.

［43］LI P，ESTRADA J L，BURLAK C，et al.Biallelic knockout of the alpha-1,3 galactosyltransferase gene in porcine liver-derived cells using zinc finger nucleases［J］.J Surg Res，2012，181(1):e39-e45.

［44］BEEVER J E.Gene-editing of porcine myostatin using zinc-finger nucleases［C］. Plant and Animal Genome XXI Conference，2013.

［45］YU S，LUO J，SONG Z，et al.Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle［J］.Cell Res，2011，21(11):1638-1640.

［46］XIONG K，LI S，ZHANG H，et al.Targeted editing of goat genome with modular-assembly zinc finger nucleases based on activity prediction by computational molecular modeling［J］.Mol Biol Rep，2013，40(7):4251-4256.

［47］PRUETT-MILLER S M，CONNELLY J P，MAEDER M L，et al.Comparison of zinc finger nucleases for use in gene targeting in mammalian cells［J］.Mol Ther，2008，16(4):707-717.

［48］CATHOMEN T，JOUNG J K.Zinc-finger nucleases:the next generation emerges［J］.Mol Ther，2008，16(7):1200-1207.

［49］FOLEY J E，YEH J R，MAEDER M L，et al.Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by Oligomerized Pool Engineering (OPEN)［J］.PLoS ONE，2009，4(2):e4348.

［50］REYON D，KIRKPATRICK J R，SANDER J D，et al.ZFNGenome:a comprehensive resource for locating zinc finger nuclease target sites in model organisms［J］.BMC Genomics，2011，12(1):83-92.

［51］MOSCOU M J，BOGDANOVE A J.A simple cipher governs DNA recognition by TAL effectors［J］.Science，2009，326(5959):1501.

［52］CHRISTIAN M，CERMAK T，DOYLE E L，et al.Targeting DNA double-strand breaks with TAL effector nucleases［J］.Genetics，2010，186(2):757-761.

［53］JOUNG J K，SANDER J D.TALENs:a widely applicable technology for targeted genome editing［J］.Nat Rev Mol Cell Biol，2013，14(1):49-55.

［54］FAHRENKRUG S C，TAN W，LILLICO S G，et al.nonmeiotic introgression of quantitative trait nucleotides and correction of congenital mutations in livestock with transcription activator-like effector nucleases［J］.Reprod Fertil Dev，2012，25(1):316.

［55］CARLSON D F，TAN W，LILLICO S G，et al.Efficient TALEN-mediated gene knockout in livestock［J］.Proc Natl Acad Sci U S A，2012，109(43):17382-17387.

［56］HUANG P，XIAO A，ZHOU M，et al.Heritable gene targeting in zebrafish using customized TALENs［J］.Nat Biotechnol, 2011，29(8):699-700.

［57］BULTMANN S，MORBITZER R，SCHMIDT C S，et al.Targeted transcriptional activation of silent oct4 pluripotency gene by combining designer TALEs and inhibition of epigenetic
modifiers［J］.Nucleic Acids Res，2012，40(12):5368-5377.

［58］CHO S W，KIM S，KIM J M，et al.Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease［J］.Nat Biotechnol，2013，31(3):230-232.