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Gene Delivery Core

The Gene Delivery Core was established in 2001 and has been supported over the years by the Program Project Grant from NHLBI. The Core's objectives are to provide investigators with access to state-of-the-art vector technology for basic research applications, to support investigators in the use of gene transfer, RNAi, protein expression and purification, and other molecular biology techniques. This objective is achieved:

  1. Through development of the new vector systems for gene delivery and gene expression.
  2. By providing molecular biology services including vector construction, virus production, and protein expression and purification.
  3. By providing the expertise to investigators, including project design and troubleshooting existing projects.
  4. By maintaining a collection of prokaryotic and eukaryotic vectors.

 

▼   Service Request

To request a service, please schedule an appointment with Director of the Gene Delivery Core, Dr. Mikhail Alexeyev. At the meeting, your gene delivery options will be discussed, including project design and choice of the vector.

Biosafety

Before service associated with generation of new recombinant DNA (rDNA) molecules can be initiated, investigators at the University of South Alabama must submit to the Institutional Biosafety Committee (IBC) a rDNA protocol, which covers construction of these molecules. The protocol must include the Director and Personnel of the Gene Delivery Core as associates. A rDNA application can be downloaded here. If you need assistance completing this form, please contact Director of the Gene Delivery Core  by calling (251) 460-6789 or emailing malexeye@southalabama.edu.

▼   Plasmid Requests
Vectors are provided either as DNA or as E. coli cultures at discretion of the Core*. The Core is committed to providing materials to investigators outside of the University of South Alabama for basic research purposes. Investigators outside University of South Alabama are requested to execute a Material Transfer Agreement (MTA). Investigators from institutions, which are signatory to UBMTA (the list of signatories to UBMTA can be found here) may execute UBMTA Implementation Letter [PDF]. Investigators from institutions not signatories to UBMTA should execute the following MTA.
▼   Core Publications
  1. Förster K, Richter H, Alexeyev MF, Rosskopf D, Felix SB, Krieg T. GSK-3beta inhibition prevents peroxide-induced collapse of mitochondrial membrane potential in rat ventricular myocytes. Clin Exp Pharmacol Physiol. 2010 Mar 5.
  2. Alexeyev MF. Is there more to aging than mitochondrial DNA and reactive oxygen species? FEBS J. 2009 Oct;276(20):5768-87.
  3. Alexeyev MF, Fayzulin R, Shokolenko IN, Pastukh V. A retro-lentiviral system for doxycycline-inducible gene expression and gene knockdown in cells with limited proliferative capacity. Mol Biol Rep. 2010 Apr;37(4):1987-91.
  4. Prasain N, Alexeyev M, Balczon R, Stevens T. Soluble adenylyl cyclase-dependent microtubule disassembly reveals a novel mechanism of endothelial cell retraction. Am J Physiol Lung Cell Mol Physiol. 2009 Jul;297(1):L73-83. Epub 2009 Apr 24.
  5. Shokolenko I, Venediktova N, Bochkareva A, Wilson GL, Alexeyev MF. Oxidative stress induces degradation of mitochondrial DNA. Nucleic Acids Res. 2009 Mar 5.
  6. Zhu B, Zhang L, Alexeyev M, Alvarez DF, Strada SJ, Stevens T. Type 5 phosphodiesterase expression is a critical determinant of the endothelial cell angiogenic phenotype. Am J Physiol Lung Cell Mol Physiol. 2009 Feb;296(2):L220-8.
  7. Lorico A, Mercapide J, Soloduschko V, Alexeyev M, Fodstad O, Rappa G. Primary neural stem/progenitor cells expressing endostatin or cytochrome P450 for gene therapy of glioblastoma. Cancer Gene Ther. 2008 Sep;15(9):605-15.
  8. Alexeyev MF, Venediktova N, Pastukh V, Shokolenko I, Bonilla G, Wilson GL. Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes. Gene Ther. 2008 Apr;15(7):516-23.
  9. Creighton J, Zhu B, Alexeyev M, Stevens T. Spectrin-anchored phosphodiesterase 4D4 restricts cAMP from disrupting microtubules and inducing endothelial cell gap formation. J Cell Sci. 2008 Jan 1;121(Pt 1):110-9.
  10. Pastukh V, Shokolenko I, Wang B, Wilson G, Alexeyev M. Human mitochondrial transcription factor A possesses multiple subcellular targeting signals. FEBS J. 2007 Dec;274(24):6488-99.
  11. Clark J, Alvarez DF, Alexeyev M, King JA, Huang L, Yoder MC, Stevens T. Regulatory role for nucleosome assembly protein-1 in the proliferative and vasculogenic phenotype of pulmonary endothelium. Am J Physiol Lung Cell Mol Physiol. 2008 Mar;294(3):L431-9.
  12. Wu S, Chen H, Alexeyev MF, King JA, Moore TM, Stevens T, Balczon RD. Microtubule motors regulate ISOC activation necessary to increase endothelial cell permeability. J Biol Chem. 2007 Nov 30;282(48):34801-8.
  13. Pastukh V, Shokolenko IN, Wilson GL, Alexeyev MF. Mutations in the passenger polypeptide can affect its partitioning between mitochondria and cytoplasm : Mutations can impair the mitochondrial import of DsRed. Mol Biol Rep. 2008 Jun;35(2):215-23.
  14. Rappa G, Anzanello F, Alexeyev M, Fodstad O, Lorico A. Gamma-glutamylcysteine synthetase-based selection strategy for gene therapy of chronic granulomatous disease and graft-vs.-host disease. Eur J Haematol. 2007 May;78(5):440-8.
  15. Zhou C, Chen H, Lu F, Sellak H, Daigle JA, Alexeyev MF, Xi Y, Ju J, van Mourik JA, Wu S. Cav3.1 (alpha1G) controls von Willebrand factor secretion in rat pulmonary microvascular endothelial cells. Am J Physiol Lung Cell Mol Physiol. 2007 Apr;292(4):L833-44.
  16. Sayner SL, Alexeyev M, Dessauer CW, Stevens T. Soluble adenylyl cyclase reveals the significance of cAMP compartmentation on pulmonary microvascular endothelial cell barrier. Circ Res. 2006 Mar 17;98(5):675-81.
  17. Alexeyev, M.F., LeDoux S.P. and Wilson, G. "Mitochondrial DNA and aging" in "Handbook of Models for Human Aging". Conn, P.M. Editor. Academic Press. 2006. PP.507-520
  18. Aronson NN Jr, Halloran BA, Alexeyev MF, Zhou XE, Wang Y, Meehan EJ, Chen L. Mutation of a conserved tryptophan in the chitin-binding cleft of Serratia marcescens chitinase A enhances transglycosylation. Biosci Biotechnol Biochem. 2006 Jan;70(1):243-51.
  19. Cioffi DL, Wu S, Alexeyev M, Goodman SR, Zhu MX, Stevens T. Activation of the endothelial store-operated ISOC Ca2+ channel requires interaction of protein 4.1 with TRPC4. Circ Res. 2005 Nov 25;97(11):1164-72.
  20. Aronson NN Jr, Halloran BA, Alexeyev MF, Zhou XE, Wang Y, Meehan EJ, Chen L. Mutation of a conserved tryptophan in the chitin-binding cleft of Serratia marcescens chitinase A enhances transglycosylation. Biosci Biotechnol Biochem. 2006 Jan;70(1):243-51.
  21. Ruchko M, Gorodnya O, Ledoux SP, Alexeyev MF, Al-Mehdi AB, Gillespie MN. Mitochondrial DNA damage triggers mitochondrial dysfunction and apoptosis in oxidant-challenged lung endothelial cells. Am J Physiol Lung Cell Mol Physiol. 2005 Mar;288(3):L530-5
  22. Shokolenko IN, Alexeyev MF, LeDoux SP, Wilson GL. TAT-mediated protein transduction and targeted delivery of fusion proteins into mitochondria of breast cancer cells. DNA Repair (Amst). 2005 Apr 4;4(4):511-8.
  23. Krieg T, Qin Q, Philipp S, Alexeyev MF, Cohen MV, Downey JM. Acetylcholine and bradykinin trigger preconditioning in the heart through a pathway that includes Akt and NOS Am J Physiol Heart Circ Physiol. 2004 Dec;287(6):H2606-11.
  24. Alexeyev MF, Ledoux SP, Wilson GL Mitochondrial DNA and aging Clin Sci (Lond). 2004 Oct;107(4):355-64
  25. Lyudmila Rachek, Valentina Grishko, Mikhail Alexeyev, Viktoriya Pastukh, Susan LeDoux, and Glenn Wilson Endonuclease III and endonuclease VIII conditionally targeted into mitochondria enhance mitochondrial DNA repair and cell survival following oxidative stress Nucleic Acids Res. 2004 Jun 15;32(10):3240-7.
  26. Alexeyev MF, Roberts RA, Daugherty RM, Audia JP, Winkler HH. Cysteine-Scanning Mutagenesis and Thiol Modification of the Rickettsia prowazekii ATP/ADP Translocase: Evidence That Transmembrane Regions I and II, but Not III, Are Structural Components of the Aqueous Translocation Channel. Biochemistry. 2004 Jun 8;43(22):6995-7002
  27. Nathan N. Aronson, Jr., Brian A. Halloran, Mikhail F. Alexeyev, Lauren Amable, Jeffry D. Madura, Lakshminarasimhulu Pasupulati, Catherine Worth and Patrick van Roey. Family 18 chitinase-oligosaccharide substrate interaction: subsite preference and anomer selectivity of Serratia marcescens chitinase A. Biochem.J. (2003) v. 376:87-95
  28. Mikhail F. Alexeyev, Viktoriya V. Pastukh, Inna N. Shokolenko, and Glenn L. Wilson. Alpha-complementation enabled T7 expression vectors and their use for the expression of recombinant polypeptides for protein transduction experiments. In: Methods in Molecular Biology. Recombinant Gene Expression:reviews and protocols. P. Balbas and A. Lorens, eds. Humana Press. 2004, 91-100.
  29. Nerino Allocati, Bartolo Favaloro, Michele Masulli, Mikhail F. Alexeyev, Carmine Di Ilio. Proteus mirabilis Glutathione S-transferase B1-1 is involved in the protective mechanisms against oxidative and chemical stresses. Biochem J. 2003 Jul 1;373(Pt 1):305-11.
  30. Inna N. Shokolenko, Mikhail F. Alexeyev, Fredica M. Robertson, Susan P. LeDoux and Glenn L. Wilson. The expression of exonuclease III from E. coli in mitochondria of breast cancer cells diminishes mitochondrial DNA repair capacity and cell survival after oxidative stress. DNA Repair (Amst). 2003 May 13;2(5):471-82.
  31. Krieg T, Landsberger M, Alexeyev MF, Felix SB, Cohen MV, Downey JM Activation of Akt is essential for acetylcholine to trigger generation of oxygen free radicals. Cardiovasc Res 2003 Apr;58(1):196-202