Engineering Enhanced Retroviral Vectors Through The Insertion Of A His6 Tag Into Vsv-g
- 详细技术说明
- None
- *Abstract
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Engineering of retroviral vectors by inserting novel protein or peptide sequences into viral proteins could readily improve their properties as gene therapy vehicles. Enhancements to VSV-G, an envelope protein commonly used to pseudotype retroviral and lentiviral vectors, could aid in vector purification and cell-specific targeting. Because the structure of VSV-G is unknown, it is difficult to rationally determine where to insert a new sequence without disrupting the virus? ability to transduce cells.
We have developed a system to modify viral genomes through the random insertion of small peptides that may confer novel function to the viral vector. This high throughput method was used to generate a library of VSV-G mutants that have a His6 tag randomly incorporated at as many or all possible points in the original protein sequence. We have screened these libraries to isolate variants that can be purified by immobilized metal affinity chromatography while still retaining their ability to transduce cells.
Purification by this method may provide advantages over current techniques in purity and scalability. In addition, these mutants will assist in identifying possible sites for the insertion of targeting ligands. The detailed genetic footprint provided by the comprehensive coverage of insertion sites will also provide further knowledge of the structure and function of VSV-G. Finally, we are utilizing this novel system to explore other desirable improvements to gene therapy vectors such as enhanced vector trafficking and antibody evasion.
- *IP Issue Date
- Apr 24, 2012
- *Principal Investigation
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Name: David Schaffer
Department:
Name: Julie Yu
Department:
- 附加资料
- Patent Number: US8163893B2
Application Number: US2008279171A
Inventor: Schaffer, David V. | Yu, Julie H.
Priority Date: 15 Feb 2006
Priority Number: US8163893B2
Application Date: 5 Feb 2009
Publication Date: 24 Apr 2012
IPC Current: C07H002104 | C12N001500 | A01N006300
US Class: 5360234 | 4353201 | 53602372
Assignee Applicant: The Regents of the University of California
Title: Pseudotyped retroviral vectors and methods of use thereof
Usefulness: Pseudotyped retroviral vectors and methods of use thereof
Summary: For production of a pseudotyped retroviral vector; useful for delivering a gene product into an individual (claimed), useful for encoding proteins for the treatment of endocrine, metabolic, hematologic, cardiovascular, neurologic, musculoskeletal, urologic, pulmonary and immune disorders; including inflammatory diseases, autoimmune, chronic and infectious diseases; such as acquired immunodeficiency syndrome (AIDS), cancer, hypercholesterolemia, insulin disorders such as diabetes, growth disorders, various blood disorders including various anemias, thalassemias and hemophilia; genetic defects such as cystic fibrosis, Gaucher's disease, Hurler's Disease, adenosine deaminase (ADA) deficiency, and emphysema.
Novelty: New nucleic acid for production of pseudotyped retroviral vector for delivering gene product comprises nucleotide sequence encoding recombinant viral envelope protein that comprises rhabdovirus envelope protein and heterologous polypeptide
- 主要类别
- 诊断/治疗
- 细分类别
- 癌症/肿瘤
- 申请号码
- 8163893
- 其他
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Tech ID/UC Case
17317/2004-010-0
Related Cases
2004-010-0
- 国家/地区
- 美国
