logging in or signing up Analysis of the HIV-1 central polypurine tract Shityakov Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT lite Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 25 Category: Science & Tech.. License: All Rights Reserved Like it (0) Dislike it (0) Added: January 16, 2012 This Presentation is Public Favorites: 0 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript Analysis of the HIV-1 central polypurine tract in a foamy viral vector background: Analysis of the HIV-1 central polypurine tract in a foamy viral vector background University of Würzburg , Institute for Virology and Immunobiology , Würzburg , Germany MD/PhD student Sergey Shityakov 8 th International Foamy Virus Conference Greece, Argos 2010PowerPoint Presentation: (-) strand DNA initiates at the primer binding site When (-) strand elongation passes the polypurine tract (ppt) region, the RNA template escapes digestion by RNase H and serves as a primer for (+) strand synthesis by DNA dependent DNA polymerization (DDDP) Reverse transcriptionPowerPoint Presentation: cPPT (HIV) vs. cPPT (PFV) GGGCAAGGGAAAGAGG GAGGAGAGGGGPowerPoint Presentation: Reverse transcription J-J Kupiec, P Sonigo, JGV 1996Aim of the project:: Aim of the project: To study the cPPT -CTS (HIV) effects upon the foamy viral replication in order to increase foamy viral vectors infectivity for growth- arrested (G1/S) permissive cells via cPPT modifications.PowerPoint Presentation: PFV and HIV systems Truncated viral backbone Packaging plasmids For PFV system : For negative control : pMD9 (Cas I, II, cPPT ) 1) pMD9 (-ENV) pGAG 2) pcDNA (mock) pPOL pENV For HIV system : pWPXL ( Ψ, Pol, cPPT-CTS ) PsPAX2 (Gag) pMD 2.G (VSV-G, pseudotyping)PowerPoint Presentation: Construction of foamy viral vectorsPowerPoint Presentation: Procedure PEI transfection of HEK 293T cells Transduction of human alveolar basal epithelial A549 cells 5μg/ml 5μg/mlPowerPoint Presentation: Transduction rates of A549 cellPowerPoint Presentation: Summary Modified foamy viral vector show a reduction in transduction rates of dividing A549 cells PFV vectors are able to infect G1/S growth-arrested A549 cells but at extremely low titers in comparison to lentiviral vectors. Finally, the HIV cPPT can not be substituted for the PFV cPPT without loss of functionality, probably because the cognate RT enzyme is required for recognition of PPT and CTS sequences.PowerPoint Presentation: Acknowledgments to PD. Dr.Carsten Scheller Axel Rethwilm’s lab: Dr.Tatiana Wiktorowicz Daniel MatthesThank you for your attention: Thank you for your attention You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.
Analysis of the HIV-1 central polypurine tract Shityakov Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT lite Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 25 Category: Science & Tech.. License: All Rights Reserved Like it (0) Dislike it (0) Added: January 16, 2012 This Presentation is Public Favorites: 0 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript Analysis of the HIV-1 central polypurine tract in a foamy viral vector background: Analysis of the HIV-1 central polypurine tract in a foamy viral vector background University of Würzburg , Institute for Virology and Immunobiology , Würzburg , Germany MD/PhD student Sergey Shityakov 8 th International Foamy Virus Conference Greece, Argos 2010PowerPoint Presentation: (-) strand DNA initiates at the primer binding site When (-) strand elongation passes the polypurine tract (ppt) region, the RNA template escapes digestion by RNase H and serves as a primer for (+) strand synthesis by DNA dependent DNA polymerization (DDDP) Reverse transcriptionPowerPoint Presentation: cPPT (HIV) vs. cPPT (PFV) GGGCAAGGGAAAGAGG GAGGAGAGGGGPowerPoint Presentation: Reverse transcription J-J Kupiec, P Sonigo, JGV 1996Aim of the project:: Aim of the project: To study the cPPT -CTS (HIV) effects upon the foamy viral replication in order to increase foamy viral vectors infectivity for growth- arrested (G1/S) permissive cells via cPPT modifications.PowerPoint Presentation: PFV and HIV systems Truncated viral backbone Packaging plasmids For PFV system : For negative control : pMD9 (Cas I, II, cPPT ) 1) pMD9 (-ENV) pGAG 2) pcDNA (mock) pPOL pENV For HIV system : pWPXL ( Ψ, Pol, cPPT-CTS ) PsPAX2 (Gag) pMD 2.G (VSV-G, pseudotyping)PowerPoint Presentation: Construction of foamy viral vectorsPowerPoint Presentation: Procedure PEI transfection of HEK 293T cells Transduction of human alveolar basal epithelial A549 cells 5μg/ml 5μg/mlPowerPoint Presentation: Transduction rates of A549 cellPowerPoint Presentation: Summary Modified foamy viral vector show a reduction in transduction rates of dividing A549 cells PFV vectors are able to infect G1/S growth-arrested A549 cells but at extremely low titers in comparison to lentiviral vectors. Finally, the HIV cPPT can not be substituted for the PFV cPPT without loss of functionality, probably because the cognate RT enzyme is required for recognition of PPT and CTS sequences.PowerPoint Presentation: Acknowledgments to PD. Dr.Carsten Scheller Axel Rethwilm’s lab: Dr.Tatiana Wiktorowicz Daniel MatthesThank you for your attention: Thank you for your attention