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Reagent Information
Name Ad-hCMV-TK 
Reagent Type Vector 
Vector
Classifications
Adenovirus
Keywords
 
Cell Line Origin  
Short Description  
Long Description This replication-defective, first generation adenoviral vector is based on human adenovirus serotype 5, with deletions in its E1a and E3 viral encoding regions. This vector encodes the transgene for Herpes Simplex Type 1 thymidine kinase under the control of the human cytomegalovirus (hCMV) promoter. The TK transgene was subcloned into the first generation adenoviral shuttle plasmid pAL119 containing the hCMV promoter to make the plasmid pAL119-TK. The adenoviral vector genome plasmid pJM17 (Microbix) and pAL119-TK were co-transfected in 293 cells (Microbix) and the adenoviral vector Ad-hCMV-TK was rescued by homologous recombination. Ad-hCMV-TK underwent three successive rounds of plaque purification and then amplified on 293 cells. Ad-hCMV-TK was purified from the 293 cell lysate by cesium chloride purification followed by three rounds of dialysis in 10mM Tris, ph 8.0.  
Vector Map Ad-hCMV-TK.doc
Sequence  
Grade Research 
Depositor Cedars-Sinai Medical Center, Los Angeles, CA 
References 1. Dewey RA, Morrissey G, Cowsill CM, Stone D, Bolognani F, Dodd NJ, Southgate TD, Klatzmann D, Lassmann H, Castro MG, Löwenstein PR. Chronic brain inflammation and persistent herpes simplex virus 1 thymidine kinase expression in survivors of syngeneic glioma treated by adenovirus-mediated gene therapy: implications for clinical trials. Nature Medicine . 1999 Nov;5(11):1256-63. PMID: 10545991. 2;Ali S, King GD, Curtin JF, Candolfi M, Xiong W, Liu C, Puntel M, Cheng Q, Prieto J, Ribas A, Kupiec-Weglinski J, van Rooijen N, Lassmann H, Lowenstein PR, Castro MG. Combined immunostimulation and conditional cytotoxic gene therapy provide long-term survival in a large glioma model. Cancer Research . 2005 Aug 15;65(16):7194-204. PMID: 16103070. 4. King GD, Muhammad AK, Curtin JF, Barcia C, Puntel M, Liu C, Honig SB, Candolfi M, Mondkar S, Lowenstein PR, Castro MG. Flt3L and TK gene therapy eradicate multifocal glioma in a syngeneic glioblastoma model . Neuro-Oncology . 2008 Feb;10(1):19-31. PMID: 18079358 5. King GD, Kroeger KM, Bresee CJ, Candolfi M, Liu C, Manalo CM, Muhammad AK, Pechnick RN, Lowenstein PR, Castro MG. Flt3L in combination with HSV1-TK-mediated gene therapy reverses brain tumor-induced behavioral deficits. Molecular Therapy. 2008 Apr;16(4):682-90. PMID: 18283279 . 6. Curtin JF, Candolfi M, Fakhouri TM, Liu C, Alden A, Edwards M, Lowenstein PR, Castro MG. Treg depletion inhibits efficacy of cancer immunotherapy: implications for clinical trials . PLoS One . 2008 Apr 23;3(4):e1983. PMID: 18431473 7. Curtin JF, Liu N, Candolfi M, Xiong W, Assi H, Yagiz K, Edwards MR, Michelsen KS, Kroeger KM, Liu C, Muhammad AK, Clark MC, Arditi M, Comin-Anduix B, Ribas A, Lowenstein PR, Castro MG. HMGB1 mediates endogenous TLR2 activation and brain tumor regression . PLoS Medicine . 2009 Jan 13;6(1):e10. PMID: 19143470 8. Candolfi M, Yagiz K, Foulad D, Alzadeh GE, Tesarfreund M, Muhammad AK, Puntel M, Kroeger KM, Liu C, Lee S, Curtin JF, King GD, Lerner J, Sato K, Mineharu Y, Xiong W, Lowenstein PR, Castro MG. Release of HMGB1 in response to proapoptotic glioma killing strategies: efficacy and neurotoxicity. Clinical Cancer Research . 2009 Jul 1;15(13):4401-14. PMID: 19570774 9. Ghulam Muhammad AK, Candolfi M, King GD, Yagiz K, Foulad D, Mineharu Y, Kroeger KM, Treuer KA, Nichols WS, Sanderson NS, Yang J, Khayznikov M, Van Rooijen N, Lowenstein PR, Castro MG. Antiglioma immunological memory in response to conditional cytotoxic/immune-stimulatory gene therapy: humoral and cellular immunity lead to tumor regression . Clinical Cancer Research . 2009 Oct 1;15(19):6113-27. PMID: 19789315. 10. Yang J, Sanderson NS, Wawrowsky K, Puntel M, Castro MG, Lowenstein PR. Kupfer-type immunological synapse characteristics do not predict anti-brain tumor cytolytic T-cell function in vivo . Proc Natl Acad Sci U S A. 2010 Mar 9;107(10):4716-21  
Biosafety The Indiana University Vector Production Facility works with this line under BL-2 containment but the level of containment required of any cell line or vector is at the discretion of the investigator's Institutional Biosafety Committee that may require a different level of containment. 
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