Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fiber-modified adenoviral vectors for enhanced transduction of tumor cells

A tumor cell and adenovirus technology, applied in the field of adenovirus vectors, can solve the problem of low efficiency of primary tumor cells

Inactive Publication Date: 2007-11-07
细胞基因系统有限公司
View PDF63 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Although adenoviral vectors have been used to transduce tumor cells, transduction of primary tumor cells is often inefficient

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fiber-modified adenoviral vectors for enhanced transduction of tumor cells
  • Fiber-modified adenoviral vectors for enhanced transduction of tumor cells
  • Fiber-modified adenoviral vectors for enhanced transduction of tumor cells

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0153] Preparation and production of adenoviral vectors

[0154] Standard systems for generating adenoviral vectors for insert expression are known in the art and are available from commercial sources, such as from Clontech (Palo Alto, CA) (Clontechniques (January 2000) pp. 10-12). Adeno-X TM Expression system, Adenovator from Qbiogene (Carlsbad, CA) TM Adenoviral vector system and AdEasy TM .

[0155]For convenience, plasmids providing the necessary parts of the adenovirus are available. Plasmid pXC.l (McKinnon (1982) Gene 19:33-42) contains the wild-type left-hand end of Ad5. PBHG10 (Bett et al. (1994); Microbix Biosystems Inc., Toronto) provides a right-hand end of Ad5 with a deletion at E3. PBHG11 provides a larger E3 deletion with an additional 0.3 kb deleted (Bett et al 1994). Alternatively, the use of pBHGE3 (Microbix Biosystems, Inc.) provides the right-hand end of Ad5 with the full length of E3.

[0156] To address early genes, the transcription start site of A...

Embodiment 1

[0250] Example 1: Human Tumor Cell Lines and Cell Culture

[0251] The human head and neck cancer lines and human melanoma cell series used in the examples described herein are listed in Table 1.

[0252] cell line

[0253] The human head and neck cancer lines and human melanoma cell lines listed in Table 1 were cultured in RPMI 1640 medium containing 10% FBS. The human prostate cancer cell line, PC3M-2AC6, was cultured in RPMI 1640 medium containing 10% FBS. The human prostate cancer cell line, LNCaP (ATCC, CRL-10995). The human prostate cancer cell line, PC-3 (ATCC, CRL-1435), was cultured in RPMI supplemented with 10% FBS and L-glutamine (2 mM).

Embodiment 2

[0254] Example 2: Construction of E1-deficient, GFP-expressing fiber chimeric Ad5 vector

[0255] To generate fiber chimera-based Ad5 vectors, the shuttle plasmid, pAd5LtRt-Smal, was first constructed as described below. First, the left-hand end of Ad5DNA (bp 1-1009bp) was amplified by PCR with two primers;

[0256] Primer 1 (5'-GAATTCTAGGGATAACAGGGTAATCATCATCAATAATACCTT-3'; SEQ ID NO: 17) and Primer 2 (5'-CCCGGGGTGCTCCACATAAATCT-3'; SEQ ID NO: 18). The right end of Ad5 DNA was amplified using a second set of primers designated primer 3 (5'-AAGCTTTAGGGATAACAGGGTAATCATCATCAATAATACCTT-3'; SEQ ID NO: 19) and primer 4 (5'-CCCGGGGGAATACATACCCGCAGG-3'; SEQ ID NO: 20) 580bp sequence. A recognition sequence for I-SceI was incorporated into primers 1 and 3. The first PCR product was digested with EcoRI and Smal and the second PCR product was digested with HindIII and Smal. The resulting fragment was gel purified and cloned by three-way ligation into the EcoRI and HindIII sites of p...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

Adenoviral vectors which effectively transduce primary tumor cells are provided. The adenoviral vectors comprise a chimeric adenovirus fiber protein which includes at least a portion of a Subgroup C adenovirus fiber shaft and at least a portion of a Subgroup B adenovirus or serotype 37 adenovirus head, wherein the head region binds CD46.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Patent Application No. 60 / 604,009, filed August 25, 2004, which is incorporated herein by reference in its entirety. Technical field: [0003] The present invention relates to adenoviral vectors comprising chimeric fibrin and exhibiting enhanced transduction of tumor cells, especially primary tumor cells, compared to non-chimeric adenoviral vectors. Background technique: [0004] Gene therapy is used as a means of delivering exogenous nucleotide sequences to cells and is the basis for some of the most novel and promising disease-fighting tools. This approach is extremely useful in the treatment of not only cancer but many other diseases including cystic fibrosis, anemia, hemophilia, diabetes, Huntington's disease, AIDS, abnormally high serum cholesterol, certain immunodeficiencies, and many forms of cancer. big prospects. Gene therapy typically relies on delivery ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12P21/06C12N15/00C12N15/83C12N15/64A61K39/12A01N63/00
Inventor 塞斯达·雷迪·鲍里斯珊希·甘斯于得超
Owner 细胞基因系统有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com