Retrograde transport viral vector system having envelope comprising fused glycoprotein
| 外国特許コード | F120007070 |
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| 整理番号 | A301-01WO |
| 掲載日 | 2012年12月10日 |
| 出願国 | 欧州特許庁（ＥＰＯ） |
| 出願番号 | 10834471 |
| 公報番号 | 2508599 |
| 公報番号 | 2508599 |
| 出願日 | 平成22年11月11日(2010.11.11) |
| 公報発行日 | 平成24年10月10日(2012.10.10) |
| 公報発行日 | 平成28年4月20日(2016.4.20) |
| 国際出願番号 | JP2010070136 |
| 国際公開番号 | WO2011068019 |
| 国際出願日 | 平成22年11月11日(2010.11.11) |
| 国際公開日 | 平成23年6月9日(2011.6.9) |
| 優先権データ |
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| 発明の名称 （英語） | Retrograde transport viral vector system having envelope comprising fused glycoprotein |
| 発明の概要（英語） |
Provided is a lentiviral vector system which sustains a high-frequency retrograde transportation ability in animal brain and has a higher titer. A kit for preparing a retrograde transport viral vector, which comprises: (1) a packaging plasmid containing the gag gene and pol gene of HIV-1; (2) a packaging plasmid containing an accessory gene of HIV-1; (3) a transfer plasmid containing a target gene; and (4) an envelope plasmid containing, as an envelope gene, a gene encoding a fused polypeptide comprising the extracellular domain of rabies virus glycoprotein (RV-G), the transmembrane domain of rabies virus glycoprotein (RV-G) or vesicular stomatitis virus glycoprotein (VSV-G) and the intracellular domain of vesicular stomatitis virus glycoprotein (VSV-G). |
| 特許請求の範囲（英語） |
[claim1] 1. A kit for preparing a retrograde transport viral vector comprising: (A) a packaging plasmid containing the gag gene and the pol gene of HIV-1; (B) a packaging plasmid containing an accessory gene of HIV-1; (C) a transfer plasmid containing a target gene; and (D) an envelope plasmid containing, as an envelope gene, a gene encoding a fused polypeptide comprising an extracellular domain of rabies virus glycoprotein (RV-G), a transmembrane domain of rabies virus glycoprotein (RV-G) or vesicular stomatitis virus glycoprotein (VSV-G) and an intracellular domain of vesicular stomatitis virus glycoprotein (VSV-G); preferably wherein the fused polypeptide has the amino acid sequence shown in SEQ ID NO 2 or SEQ ID NO 6, and the packaging plasmid (1) is pCAGkGP1.1 R, and the packaging plasmid (2) is pCAG4-RTR2, and the transfer plasmid is pCL20c-MSCV-X where "X" represents the target gene; more preferably wherein the envelope gene is expressed under control of a cytomegalovirus enhancer and an avian beta -actin promoter in the envelope plasmid; even more preferably wherein a base sequence of the gene encoding the fused polypeptide is shown in SEQ ID NO 1 or SEQ ID NO 5. [claim2] 2. The kit for preparing a viral vector according to Claim 1, wherein the target gene is a human gene, preferably wherein the target gene is a gene for treating a cranial nerve disease. [claim3] 3. A kit for producing a producer cell, comprising the kit for preparing a viral vector according to Claims 1 or 2 and a host cell, preferably wherein an infected cell is a HEK293 T-cell. [claim4] 4. A method of producing a producer cell, comprising co-transfecting the infected cell with the packaging plasmid, the transfer plasmid and the envelope plasmid contained in the kit for preparing a viral vector according to Claims 1 or 2; preferably wherein the infected cell is a HEK293 T-cell; more preferably wherein the transfection is performed using the calcium phosphate method. [claim5] 5. The producer cell obtained by the method according to Claim 4. [claim6] 6. A method of producing a viral vector, comprising: culturing the producer cell according to Claim 5 and harvesting a virus particle from the supernatant of the culture. [claim7] 7. A viral vector possessing a retrograde transportation ability, produced by the method according to Claim 6. [claim8] 8. A viral vector as claimed in claim 7 for use in a method of gene transfer, the method comprising: infecting a nerve terminal of an animal with the viral vector; introducing the viral vector into a cell body of the nerve at a target region in a brain by retrograde transportation of the viral vector through an axon of the nerve; and expressing the target gene in the cell body; preferably wherein the nerve terminal is located in striatum, and the target region in the brain is a region in the brain center which projects to striatum; more preferably wherein: (a) the region in the brain center which projects to striatum is primary motor cortex, primary somatosensory cortex, parafascicular nucleus of thalamus and/or substantia nigra pars compacta; or (b) the region in the brain center which projects to ventral striatum (nucleus accumbens) is piriform cortex, subiculum, amygdala basolateral nucleus, anterior paraventricular nucleus, mediodorsal nucleus of thalamus and/or lateral hypothalamus. [claim9] 9. A viral vector as claimed in Claim 8, wherein the animal is a mammal, preferably wherein the mammal is a primate, more preferably wherein the primate is a human. [claim10] 10. An agent for gene therapy, containing the viral vector according to Claim 7 as an active ingredient. [claim11] 11. A viral vector as claimed in claim 8 or 9, wherein the target gene introduced by the method of gene transfer is integrated into the chromosome of a cell at the target region to be expressed, and wherein the method is a method of gene therapy for a brain disease, preferably wherein the brain disease is Parkinson's disease. [claim12] 12. A viral vector as claimed in claim 11, wherein the method of gene therapy comprises administering the agent for gene therapy according to Claim 10 to a patient. [claim13] 13. An envelope for pseudotyping a lentiviral vector, comprising a fused polypeptide comprising an extracellular domain and a transmembrane domain of rabies virus glycoprotein (RV-G), and an intracellular domain of vesicular stomatitis virus glycoprotein (VSV-G), preferably wherein the lentiviral vector is HIV-1 lentivirus. [claim14] 14. A gene encoding the envelope comprising the fused polypeptide according to Claim 13. [claim15] 15. An envelope plasmid containing the gene according to Claim 14. |
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| 国際特許分類(IPC) |
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| 参考情報 （研究プロジェクト等） | CREST 脳の機能発達と学習メカニズムの解明 領域 |
日本語項目の表示
| 発明の名称 | 融合糖タンパク質から成るエンベロープを有する逆行性輸送ウイルスベクター系 |
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