Name: production of lentiviral vectors for transducing cells from the central nervous system
Uploaded: 2012-05-24T13:00:00
Description: Watch this Scientific Journal Video about Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System at JoVE.com

Эта работа была поддержана NIH Neuroscience Blueprint Основные гранта (P30 NS057105, BJS) в Вашингтонском университете, Программа гранта NS032636 (BJS) и Надежда Центра неврологических расстройств.

1. Упаковка лентивирусов векторов Лентивирусов векторов производятся котрансфекции из лентивирусов вектор передачи и другие плазмиды, необходимые для упаковки в 293T клеток кальцием метод трансфекции фосфатом. Мы используем 10 100-мм культуры тканей блюда в этом протоколе. ...

<ol>
	<li>Naldini, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+gene+delivery+and+stable+transduction+of+nondividing+cells+by+a+lentiviral+vector.">In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector.</a> Science. 272, 263-267 (1996).</li><li>Zufferey, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-inactivating+lentivirus+vector+for+safe+and+efficient+in+vivo+gene+delivery.">Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery.</a> J. Virol. 72, 9873-9880 (1998).</li><li>Davidson, B. L., Breakefield, X. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+vectors+for+gene+delivery+to+the+nervous+system.">Viral vectors for gene delivery to the nervous system.</a> Nat. Rev. Neurosci. 4, 353-364 (2003).</li><li>Gascon, S., Paez-Gomez, J. A., Diaz-Guerra, M., Scheiffele, P., Scholl, F. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual-promoter+lentiviral+vectors+for+constitutive+and+regulated+gene+expression+in+neurons.">Dual-promoter lentiviral vectors for constitutive and regulated gene expression in neurons.</a> J. Neurosci. Methods. 168, 104-1012 (2008).</li><li>Hioki, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+transduction+of+neurons+by+lentivirus+with+enhanced+neuron-specific+promoters.">Efficient gene transduction of neurons by lentivirus with enhanced neuron-specific promoters.</a> Gene Ther. 14, 872-882 (2007).</li><li>Li, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+promoter+usage+for+lentiviral+vector-mediated+transduction+of+cultured+central+nervous+system+cells.">Optimal promoter usage for lentiviral vector-mediated transduction of cultured central nervous system cells.</a> J. Neurosci. Methods. 189, 56-64 (2010).</li><li>Naldini, L., Blomer, U., Gage, F. H., Trono, D., Verma, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+transfer,+integration,+and+sustained+long-term+expression+of+the+transgene+in+adult+rat+brains+injected+with+a+lentiviral+vector.">Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector.</a> Proc. Natl. Acad. Sci. U.S.A. 93, 11382-11388 (1996).</li><li>Blomer, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+efficient+and+sustained+gene+transfer+in+adult+neurons+with+a+lentivirus+vector.">Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector.</a> J. Virol. 71, 6641-6649 (1997).</li><li>Consiglio, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Robust+in+vivo+gene+transfer+into+adult+mammalian+neural+stem+cells+by+lentiviral+vectors.">Robust in vivo gene transfer into adult mammalian neural stem cells by lentiviral vectors.</a> Proc. Natl. Acad. Sci. U.S.A. 101, 14835-14840 (2004).</li><li>Jakobsson, J., Ericson, C., Jansson, M., Bjork, E., Lundberg, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+transgene+expression+in+rat+brain+using+lentiviral+vectors.">Targeted transgene expression in rat brain using lentiviral vectors.</a> J. Neurosci. Res. 73, 876-885 (2003).</li><li>Arya, S. K., Guo, C., Josephs, S. F., Wong-Staal, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trans-activator+gene+of+human+T-lymphotropic+virus+type+III+(HTLV-III).">Trans-activator gene of human T-lymphotropic virus type III (HTLV-III).</a> Science. 229, 69-73 (1985).</li><li>Sirven, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+human+immunodeficiency+virus+type-1+central+DNA+flap+is+a+crucial+determinant+for+lentiviral+vector+nuclear+import+and+gene+transduction+of+human+hematopoietic+stem+cells.">The human immunodeficiency virus type-1 central DNA flap is a crucial determinant for lentiviral vector nuclear import and gene transduction of human hematopoietic stem cells.</a> Blood. 96, 4103-4110 (2000).</li><li>Burns, J. C., Friedmann, T., Driever, W., Burrascano, M., Yee, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vesicular+stomatitis+virus+G+glycoprotein+pseudotyped+retroviral+vectors:+concentration+to+very+high+titer+and+efficient+gene+transfer+into+mammalian+and+nonmammalian+cells.">Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells.</a> Proc. Natl. Acad. Sci. U.S.A. 90, 8033-8037 (1993).</li><li>Farson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new-generation+stable+inducible+packaging+cell+line+for+lentiviral+vectors.">A new-generation stable inducible packaging cell line for lentiviral vectors.</a> Hum. Gene Ther. 12, 981-997 (2001).</li><li>Broussau, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducible+packaging+cells+for+large-scale+production+of+lentiviral+vectors+in+serum-free+suspension+culture.">Inducible packaging cells for large-scale production of lentiviral vectors in serum-free suspension culture.</a> Mol. Ther. 16, 500-507 (2008).</li><li>Wang, X., McManus, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lentivirus+production.">Lentivirus production.</a> J. Vis. Exp. (32), e1499 (2009).</li><li>Sastry, L., Johnson, T., Hobson, M. J., Smucker, B., Cornetta, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Titering+lentiviral+vectors:+comparison+of+DNA,+RNA+and+marker+expression+methods.">Titering lentiviral vectors: comparison of DNA, RNA and marker expression methods.</a> Gene Ther. 9, 1155-1162 (2002).</li><li>Snider, B. J., Lobner, D., Yamada, K. A., Choi, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conditioning+heat+stress+reduces+excitotoxic+and+apoptotic+components+of+oxygen-glucose+deprivation-induced+neuronal+death+in+vitro.">Conditioning heat stress reduces excitotoxic and apoptotic components of oxygen-glucose deprivation-induced neuronal death in vitro.</a> J. Neurochem. 70, 120-129 (1998).</li><li>Mazarakis, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rabies+virus+glycoprotein+pseudotyping+of+lentiviral+vectors+enables+retrograde+axonal+transport+and+access+to+the+nervous+system+after+peripheral+delivery.">Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery.</a> Hum. Mol. Genet. 10, 2109-2121 (2001).</li><li>Kato, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuron-specific+gene+transfer+through+retrograde+transport+of+lentiviral+vector+pseudotyped+with+a+novel+type+of+fusion+envelope+glycoprotein.">Neuron-specific gene transfer through retrograde transport of lentiviral vector pseudotyped with a novel type of fusion envelope glycoprotein.</a> Hum. Gene Ther. 22, 1511-1523 (2011).</li><li>Segura, M. M., Garnier, A., Durocher, Y., Ansorge, S., Kamen, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+protocol+for+lentiviral+vector+mass+production.">New protocol for lentiviral vector mass production.</a> Methods Mol. Biol. 614, 39-52 (2010).</li><li>Kutner, R. H., Puthli, S., Marino, M. P., Reiser, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+production+and+concentration+of+HIV-1-based+lentiviral+vectors+using+HYPERFlask+vessels+and+anion+exchange+membrane+chromatography.">Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography.</a> BMC Biotechnol. 9, 10 (2009).</li><li>Laughlin, M. A., Chang, G. Y., Oakes, J. W., Gonzalez-Scarano, F., Pomerantz, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sodium+butyrate+stimulation+of+HIV-1+gene+expression:+a+novel+mechanism+of+induction+independent+of+NF-kappa+B.">Sodium butyrate stimulation of HIV-1 gene expression: a novel mechanism of induction independent of NF-kappa B.</a> J Acquir Immune Defic Syndr Hum Retrovirol. 9, 332-339 (1995).</li><li>Gasmi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirements+for+efficient+production+and+transduction+of+human+immunodeficiency+virus+type+1-based+vectors.">Requirements for efficient production and transduction of human immunodeficiency virus type 1-based vectors.</a> J. Virol. 73, 1828-1834 (1999).</li><li>Palsson, B., Andreadis, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physico-chemical+factors+that+govern+retrovirus-mediated+gene+transfer.">The physico-chemical factors that govern retrovirus-mediated gene transfer.</a> Exp. Hematol. 25, 94-102 (1997).</li><li>Lizee, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+quantitative+reverse+transcriptase-polymerase+chain+reaction+as+a+method+for+determining+lentiviral+vector+titers+and+measuring+transgene+expression.">Real-time quantitative reverse transcriptase-polymerase chain reaction as a method for determining lentiviral vector titers and measuring transgene expression.</a> Hum. Gene Ther. 14, 497-507 (2003).</li><li>Lee, J. K., Chung, J., McAlpine, F. E., Tansey, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulator+of+G-Protein+Signaling-10+Negatively+Regulates+NF-{kappa}B+in+Microglia+and+Neuroprotects+Dopaminergic+Neurons+in+Hemiparkinsonian+Rats.">Regulator of G-Protein Signaling-10 Negatively Regulates NF-{kappa}B in Microglia and Neuroprotects Dopaminergic Neurons in Hemiparkinsonian Rats.</a> J. Neurosci. 31, 11879-11888 (2011).</li><li>Shevtsova, Z., Malik, J. M., Michel, U., Bahr, M., Kugler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Promoters+and+serotypes:+targeting+of+adeno-associated+virus+vectors+for+gene+transfer+in+the+rat+central+nervous+system+in+vitro+and+in+vivo.">Promoters and serotypes: targeting of adeno-associated virus vectors for gene transfer in the rat central nervous system in vitro and in vivo.</a> Exp. Physiol. 90, 53-59 (2005).</li></ol>

В этом протоколе, мы показали, производство лентивирусов векторов и применение этих векторов в неокортекса культур. Мы продемонстрировали эффективную и клетки определенного типа трансдукции с векторами производятся с помощью этих методов. Когда промоутер synapsin использовании GFP выраж?...

<table border="1"><tbody><tr><td> Название реагента </td><td> Компания </td><td> Номер по каталогу </td></tr><tr><td> DMEM </td><td> Sigma-Aldrich </td><td> D5796 </td></tr><tr><td> MEM </td><td> Invitrogen </td><td> 11090-081 </td></tr><tr><td> Эмбриональной телячьей сыворотки </td><td> Hyclone </td><td> SV3001403 </td></tr><tr><td> PBS </td><td> Mediatech </td><td> 21-040-СМ </td></tr><tr><td> Трипсин-EDTA </td><td> Sigma-Aldrich </td><td> T3924 </td></tr><tr><td> Бутират натрия </td><td> Sigma-Aldrich </td><td> B5887 </td></tr><tr><td> Hexadimethrine бромид (Polybrene) </td><td> Sigma-Aldrich </td><td> H9268 </td></tr><tr><td> 293T клеток </td><td> ATCC </td><td> CRL-11268 </td></tr><tr><td> HT1080 клеток </td><td> ATCC </td><td> CCL-121 </td></tr><tr><td> Сокол 100 х 20 мм ткань культурноэлектронной блюдо </td><td> BD Biosciences </td><td> 353003 </td></tr><tr><td> 1 х 3 ½ в трубы центрифуги polyallomoer </td><td> Beckman Coulter- </td><td> 326823 </td></tr><tr><td> 0,2-микронной шприц фильтр </td><td> Гранулирование </td><td> 431219 </td></tr><tr><td> QIAamp ДНК Mini Kit </td><td> Qiagen </td><td> 51304 </td></tr></tbody></table>

production of lentiviral vectors for transducing cells from the central nervous system

Efficient gene delivery in the central nervous system (CNS) is important in studying gene functions, modeling neurological diseases and developing therapeutic approaches. Lentiviral vectors are attractive tools in transduction of neurons and other cell types in CNS as they transduce both dividing and non-dividing cells, support sustained expression of transgenes, and have relatively large packaging capacity and low toxicity 1-3. Lentiviral vectors have been successfully used in transducing many neural cell types in vitro 4-6 and in animals 7-10.
Great efforts have been made to develop lentiviral vectors with improved biosafety and efficiency for gene delivery. The current third generation replication-defective and self-inactivating (SIN) lentiviral vectors are depicted in Figure 1. The required elements for vector packaging are split into four plasmids. In the lentiviral transfer plasmid, the U3 region in the 5' long terminal repeat (LTR) is replaced with a strong promoter from another virus. This modification allows the transcription of the vector sequence independent of HIV-1 Tat protein that is normally required for HIV gene expression 11. The packaging signal (&Psi;) is essential for encapsidation and the Rev-responsive element (RRE) is required for producing high titer vectors. The central polypurine tract (cPPT) is important for nuclear import of the vector DNA, a feature required for transducing non-dividing cells 12. In the 3' LTR, the cis-regulatory sequences are completely removed from the U3 region. This deletion is copied to 5' LTR after reverse transcription, resulting in transcriptional inactivation of both LTRs. Plasmid pMDLg/pRRE contains HIV-1 gag/pol genes, which provide structural proteins and reverse transcriptase. pRSV-Rev encodes Rev which binds to the RRE for efficient RNA export from the nucleus. pCMV-G encodes the vesicular stomatitis virus glycoprotein (VSV-G) that replaces HIV-1 Env. VSV-G expands the tropism of the vectors and allows concentration via ultracentrifugation 13. All the genes encoding the accessory proteins, including Vif, Vpr, Vpu, and Nef are excluded in the packaging system. The production and manipulation of lentiviral vectors should be carried out according to NIH guidelines for research involving recombinant DNA (<a href="http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.pdf" target="_blank">http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.pdf</a>). An approval from individual Institutional Biological and Chemical Safety Committee may be required before using lentiviral vectors. Lentiviral vectors are commonly produced by cotransfection of 293T cells with lentiviral transfer plasmid and the helper plasmids encoding the proteins required for vector packaging. Many lentiviral transfer plasmids and helper plasmids can be obtained from Addgene, a non-profit plasmid repository (<a href="http://www.addgene.org/" target="_blank">http://www.addgene.org/</a>). Some stable packaging cell lines have been developed, but these systems provide less flexibility and their packaging efficiency generally declines over time 14, 15. Commercially available transfection kits may support high efficiency of transfection 16, but they can be very expensive for large scale vector preparations. Calcium phosphate precipitation methods provide highly efficient transfection of 293T cells and thus provide a reliable and cost effective approach for lentiviral vector production.
In this protocol, we produce lentiviral vectors by cotransfection of 293T cells with four plasmids based on the calcium phosphate precipitation principle, followed by purification and concentration with ultracentrifugation through a 20% sucrose cushion. The vector titers are determined by fluorescence- activated cell sorting (FACS) analysis or by real time qPCR. The production and titration of lentiviral vectors in this protocol can be finished with 9 days. We provide an example of transducing these vectors into murine neocortical cultures containing both neurons and astrocytes. We demonstrate that lentiviral vectors support high efficiency of transduction and cell type-specific gene expression in primary cultured cells from CNS.

Watch this Scientific Journal Video about Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System at JoVE.com

Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System

В этом протоколе описываются производства, очистки и титрования лентивирусов векторов. Мы приведем пример того, лентивирусов вектор-опосредованной доставки генов в первичных культивируемых нейронов и астроцитов. Наши методы могут применяться и к другим типам клеток<em&gt; В пробирке</em&gt; И<em&gt; В естественных условиях</em&gt;.

Производство лентивирусов векторов для трансдукции клетки из центральной нервной системы

Efficient gene delivery in the central nervous system (CNS) is important in studying gene functions, modeling neurological diseases and developing ...

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