Team:Freiburg Bioware/Filelist
From 2010.igem.org
VolkerMorath (Talk | contribs) |
|||
(48 intermediate revisions not shown) | |||
Line 7: | Line 7: | ||
</html> | </html> | ||
- | |||
- | |||
- | + | Finally we successfully combined different modifactions of the therapeutic viral vector for optimizing it for tumore therapy application. | |
+ | These modifications include the knock down of the natural tropism for HSPG receptor, the tumor targeting using the affibody ZEGFR1907 fused to the N-terminus of the viral coat protein VP2 and an encapsidated vectorgenome that was bricked down and reassembled containing the prodrug converting Guanosine Monophosphate Kinase Thymidine Kinase fusion protein (mGMK-TK). | ||
- | |||
- | |||
- | [[Media: | + | <h2>AAAAAH</h2> |
+ | [[Media:Freiburg10_Organisation_cap_proteins.png]] | ||
+ | [[Media:Freiburg10 Organisation rep proteins.png]] | ||
- | [[Media: | + | [[Media:Freiburg10_Freiburg10_amidosulfated_disacharid.png]] |
+ | [[Media:Freiburg10 Organisation rep proteins.png]] | ||
- | |||
- | [[Media: | + | <h2>eGFP</h2> |
+ | [[Media:Freiburg10_eGFP.png]] | ||
+ | [[Media:Freiburg10_CD.png]] | ||
+ | [[Media:Freiburg10_pathway_CD.png]] | ||
+ | [[Media:Freiburg10_CFP_spectrum]] | ||
+ | [[Media:Freiburg10_CFP_fluorophor]] | ||
+ | [[Media:Freiburg10_CFP_architecture]] | ||
- | |||
- | |||
- | [[Media: | + | [[Media:Freiburg10_Loopover1.jpeg]] |
- | [[Media: | + | [[Media:Freiburg10_Loopover2.jpeg]] |
- | + | <h2>Time lapse</h2> | |
- | [[Media: | + | [[Media:Freiburg10_production_Venus_mCherry_Brightfield.mov]] |
- | [[Media: | + | [[Media:Freiburg10_TimeLapse_HT10180_TK_GMK_400px.gif]] |
- | [[Media: | + | [[Media:Freiburg10_TimeLapse_A431_TK_GMK_400px.gif]] |
- | [[Media: | + | [[Media:Freiburg10_TimeLapse_AAV-293_mVenus+GOI_mCherry_390px.gif]] |
- | [[Media: | + | [[Media:Freiburg10_TimeLapse_AAV-293_mVenus+GOI_mCherry_400px.gif]] |
+ | |||
+ | |||
+ | <h2>Structure Modeling</h2> | ||
[[Media:Freiburg10_rot250trans.gif]] | [[Media:Freiburg10_rot250trans.gif]] | ||
Line 51: | Line 57: | ||
[[Media:Freiburg10_AAVassem500-100.mov]] | [[Media:Freiburg10_AAVassem500-100.mov]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_complete.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_singleVP3.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_singleVP3surface.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_VP3star.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_slice.gif]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_sliceSur.gif]] | ||
+ | |||
+ | [[Media:Freiburg10_PleaseWait.gif]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_assembly_600px.png]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_assembly_400px.png]] | ||
+ | |||
+ | [[Media:Freiburg10_AAV_assembly_scheme_400px.png]] | ||
+ | |||
<h2>Modularaization of GOI - Results</h2> | <h2>Modularaization of GOI - Results</h2> | ||
Line 64: | Line 91: | ||
[[Media:Freiburg10_Cloning_Full_GOI.png]] | [[Media:Freiburg10_Cloning_Full_GOI.png]] | ||
- | [[Media: | + | [[Media:Freiburg10_Microscopy_Overlay_mVenus_1_GOI.png]] |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | [[Media: | + | [[Media:Freiburg10_Microscopy_Overlay_grey_1_GOI.png]] |
- | [[Media: | + | [[Media:Freiburg10_Microscopy_Overlay_mVenus_2_GOI.png]] |
- | [[Media: | + | [[Media:Freiburg10_Microscopy_phasecontrast_2_GOI.png]] |
- | + | ||
- | + | ||
[[Media:Freiburg10_FACS_woHGH.png]] | [[Media:Freiburg10_FACS_woHGH.png]] | ||
Line 100: | Line 117: | ||
[[Media:Freiburg10_FACS_FULL_pAAV_mVenus.png]] | [[Media:Freiburg10_FACS_FULL_pAAV_mVenus.png]] | ||
- | [[Media:Freiburg10_Diagram_FULL_GOI.png]] | + | [[Media:Freiburg10_Diagram_FULL_GOI.png]] **neues Bild hochladen!** |
Line 123: | Line 140: | ||
[[Media:Freiburg10_Crystal_structure_SF3helicase.png]] | [[Media:Freiburg10_Crystal_structure_SF3helicase.png]] | ||
+ | |||
+ | [[Media:Freiburg10_PLA2-mechanism.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10_PLA2-recognition_site.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_VP1-3_overview.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_Rep40.jpg]] | ||
<h2> Trafficking </h2> | <h2> Trafficking </h2> | ||
Line 148: | Line 173: | ||
[[Media:Freiburg10_Flourocytosine_pathway.png]] | [[Media:Freiburg10_Flourocytosine_pathway.png]] | ||
+ | |||
+ | [[Media:Freiburg10_CD.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_TK.jpg]] | ||
<h2> Loop insertions </h2> | <h2> Loop insertions </h2> | ||
Line 154: | Line 183: | ||
[[Media:Freiburg10_cloning_viral_bricks.png]] | [[Media:Freiburg10_cloning_viral_bricks.png]] | ||
+ | |||
+ | [[Media:Freiburg10_ELISA.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_ELISA_plate.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_akta.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_his_column.jpg]] | ||
+ | |||
+ | [[Media:Freiburg10_His-tag.png]] | ||
+ | |||
+ | [[Media:Freiburg10_His-experiment.png]] | ||
+ | |||
+ | [[Media:Freiburg10_His-data.png]] | ||
+ | |||
+ | [[Media:Freiburg10_BAP-tag.png]] | ||
+ | |||
+ | [[Media:Freiburg10_BAP-experiment.png]] | ||
+ | |||
+ | [[Media:Freiburg10_BAP-data.png]] | ||
+ | |||
+ | [[Media:Freiburg10_Z34C-tag.png]] | ||
+ | |||
+ | [[Media:Freiburg10_Z34C-history.png]] | ||
+ | |||
+ | [[Media:Freiburg10_Z34C affinity data.png]] | ||
+ | |||
+ | [[Media:Freiburg10_Z34C titration data.png]] | ||
+ | |||
+ | [[Media:Freiburg10_loop insertion sites.png]] | ||
+ | |||
+ | |||
Line 176: | Line 237: | ||
<h2> N-terminal fusion </h2> | <h2> N-terminal fusion </h2> | ||
+ | ===Results=== | ||
+ | [[Media:Freiburg10 mVenusDrop.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 qPCR DARPin.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS1.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS2.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS3.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 Cloning VP2Fusion2.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 Cloning VP2FusionH.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 CloningGel DARPin.png]] | ||
+ | |||
+ | [[Media:Freiburg10 CMCSchema.png]] | ||
+ | |||
+ | [[Media:Freiburg10 ColonyPCR.png]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS VP1Ins A431.png]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS VP1Ins HT1080.png]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS VP2 A431.png]] | ||
+ | |||
+ | [[Media:Freiburg10 FACS VP2Fusion HT1080 Diagramm.png]] | ||
+ | |||
+ | [[Media:Freiburg10 Gel VP1ins.png]] | ||
+ | |||
+ | [[Media:Freiburg10 GelVP2Fusion2tog.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 GelVP2Fusion2tog2.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 LinkerSchema.png]] | ||
+ | |||
+ | [[Media:Freiburg10 NLS mVenus mCherry.png]] | ||
+ | |||
+ | [[Media:Freiburg10 NLSSchema.png]] | ||
+ | |||
+ | [[Media:Freiburg10 Overview VP1Insertion.png]] | ||
+ | |||
+ | [[Media:Freiburg10 Overview VP2Fusion.png]] | ||
+ | |||
+ | [[Media:Freiburg10 Primer ColonyPCR.png]] | ||
+ | |||
+ | [[Media:Freiburg10 qPCR VP1Insnew.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 qPCR VP2Fus.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 SchemaVP2Fusion.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 TargetingMotifSchema.png]] | ||
+ | |||
+ | [[Media:Freiburg10 test supernatant pellet.png]] | ||
+ | |||
+ | [[Media:Freiburg10 TestDARPin.png]] | ||
+ | |||
+ | [[Media:Freiburg10 TimeLapseA431.png]] | ||
+ | |||
+ | [[Media:Freiburg10 TimeLapseHT1080.png]] | ||
+ | |||
+ | [[Media:Freiburg10 VP1InsertionSchema.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 VP1KO.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 VP1upSchema.png]] | ||
+ | |||
+ | [[Media:Freiburg10 VP2KO.PNG]] | ||
+ | |||
+ | [[Media:Freiburg10 VP23Schema.png]] | ||
+ | |||
[[Media:Freiburg10_N_terminal_fusion_approach.png]] | [[Media:Freiburg10_N_terminal_fusion_approach.png]] | ||
Line 217: | Line 351: | ||
[[Media:Freiburg10_Darpin_capping_modules.png]] | [[Media:Freiburg10_Darpin_capping_modules.png]] | ||
+ | |||
+ | <h2> HSPG KO </h2> | ||
+ | |||
+ | [[Media:Freiburg10 HSPG binding motif.png]] | ||
+ | |||
+ | [[Media:Freiburg10 heparansulfate.PNG]] | ||
+ | |||
+ | <h2>ITR Diary </h2> | ||
+ | |||
+ | |||
+ | <h2> Targeting: Flow Cytometry: DARPin, Affi (HSPGKO) </h2> | ||
+ | |||
+ | |||
+ | [[Media:Freiburg10 FACS DARPin Affi HSPGKO A431.png]] | ||
+ | |||
+ | [[Media:Freiburg10_FACS DARPin Affi HSPGKO HT1080.png]] | ||
+ | |||
+ | [[Media:Freiburg10_FlowCytometry Diagramm DARPin Affi.png]] |
Latest revision as of 03:49, 28 October 2010
File list
Finally we successfully combined different modifactions of the therapeutic viral vector for optimizing it for tumore therapy application.
These modifications include the knock down of the natural tropism for HSPG receptor, the tumor targeting using the affibody ZEGFR1907 fused to the N-terminus of the viral coat protein VP2 and an encapsidated vectorgenome that was bricked down and reassembled containing the prodrug converting Guanosine Monophosphate Kinase Thymidine Kinase fusion protein (mGMK-TK).
AAAAAH
Media:Freiburg10_Organisation_cap_proteins.png Media:Freiburg10 Organisation rep proteins.png
Media:Freiburg10_Freiburg10_amidosulfated_disacharid.png Media:Freiburg10 Organisation rep proteins.png
eGFP
Media:Freiburg10_eGFP.png Media:Freiburg10_CD.png Media:Freiburg10_pathway_CD.png Media:Freiburg10_CFP_spectrum Media:Freiburg10_CFP_fluorophor Media:Freiburg10_CFP_architecture
Media:Freiburg10_Loopover1.jpeg
Media:Freiburg10_Loopover2.jpeg
Time lapse
Media:Freiburg10_production_Venus_mCherry_Brightfield.mov
Media:Freiburg10_TimeLapse_HT10180_TK_GMK_400px.gif
Media:Freiburg10_TimeLapse_A431_TK_GMK_400px.gif
Media:Freiburg10_TimeLapse_AAV-293_mVenus+GOI_mCherry_390px.gif
Media:Freiburg10_TimeLapse_AAV-293_mVenus+GOI_mCherry_400px.gif
Structure Modeling
Media:Freiburg10_rot250trans.gif
Media:Freiburg10_AAVassem500-60.gif
Media:Freiburg10_AAVassem500-100.gif
Media:Freiburg10_AAVassem500-100.mov
Media:Freiburg10_AAV_complete.jpg
Media:Freiburg10_AAV_singleVP3.jpg
Media:Freiburg10_AAV_singleVP3surface.jpg
Media:Freiburg10_AAV_VP3star.jpg
Media:Freiburg10_AAV_slice.gif
Media:Freiburg10_AAV_sliceSur.gif
Media:Freiburg10_PleaseWait.gif
Media:Freiburg10_AAV_assembly_600px.png
Media:Freiburg10_AAV_assembly_400px.png
Media:Freiburg10_AAV_assembly_scheme_400px.png
Modularaization of GOI - Results
Media:Freiburg10_Overview_BBa_Vectorplasmid.png
Media:Freiburg10_Overview_Assembly_GOI.png
Media:Freiburg10_TheoreticalCloning_ITR_CMV_beta.png
Media:Freiburg10_Cloning_Intermediate_GOI.png
Media:Freiburg10_Cloning_Full_GOI.png
Media:Freiburg10_Microscopy_Overlay_mVenus_1_GOI.png
Media:Freiburg10_Microscopy_Overlay_grey_1_GOI.png
Media:Freiburg10_Microscopy_Overlay_mVenus_2_GOI.png
Media:Freiburg10_Microscopy_phasecontrast_2_GOI.png
Media:Freiburg10_FACS_woHGH.png
Media:Freiburg10_FACS_withHGH.png
Media:Freiburg10_Diagram_hGH.png
Media:Freiburg10_FACS_betaglobin.png
Media:Freiburg10_FACS_withbetaglobin.png
Media:Freiburg10 VirusInfectionScheme01.png
Media:Freiburg10_Diagram_betaglobin.png.png
Media:Freiburg10_FACS_FULL_pSB1c3_mVenus.png
Media:Freiburg10_FACS_FULL_pAAV_mVenus.png
Media:Freiburg10_Diagram_FULL_GOI.png **neues Bild hochladen!**
Introduction to Adeno-Associated Virus Serotype 2
Media:Freiburg10_genomic_organisation_wt.png
Media:Freiburg10_organization_ITRs.png
Media:Freiburg10_Overview_AAV-2_replication.png
Media:Freiburg10_regulation_viral_promotors.png
Media:Freiburg10_p5_promoter_wtAAV2.png
Media:Freiburg10_p5_TATA_less.png
Media:Freiburg10_DNA_Loop_P5_to_sp1.png
Media:Freiburg10_Rep_proteins_organization.png
Media:Freiburg10_Crystal_structure_SF3helicase.png
Media:Freiburg10_PLA2-mechanism.PNG
Media:Freiburg10_PLA2-recognition_site.jpg
Media:Freiburg10_VP1-3_overview.jpg
Trafficking
Media:Freiburg10_viral_trafficking.png
Media:Freiburg10_cellular_surface_HSPG.png
Media:Freiburg10_Clathrin_triskelions.png
Helper genes
Media:Freiburg10_Helper_free_system.png
Gene therapy
Media:Freiburg10_overview_GDEPT.png
Media:Freiburg10_Bystander_effect.png
Cytosine deaminase
Media:Freiburg10_Flourocytosine_pathway.png
Loop insertions
Media:Freiburg10_3D_AC_of_453_587_loops.png
Media:Freiburg10_cloning_viral_bricks.png
Media:Freiburg10_ELISA_plate.jpg
Media:Freiburg10_his_column.jpg
Media:Freiburg10_His-experiment.png
Media:Freiburg10_BAP-experiment.png
Media:Freiburg10_Z34C-history.png
Media:Freiburg10_Z34C affinity data.png
Media:Freiburg10_Z34C titration data.png
Media:Freiburg10_loop insertion sites.png
Tagging
Targeting
Media:Freiburg10_RGD_motif.png
Media:Freiburg10_Z34C_motif.png
Media:Freiburg10_Z34C_Z38_ZDomain_helix.png
Media:Freiburg10_HSPG_chemical_structure.png
N-terminal fusion
Results
Media:Freiburg10 mVenusDrop.PNG
Media:Freiburg10 qPCR DARPin.PNG
Media:Freiburg10 Cloning VP2Fusion2.PNG
Media:Freiburg10 Cloning VP2FusionH.PNG
Media:Freiburg10 CloningGel DARPin.png
Media:Freiburg10 CMCSchema.png
Media:Freiburg10 ColonyPCR.png
Media:Freiburg10 FACS VP1Ins A431.png
Media:Freiburg10 FACS VP1Ins HT1080.png
Media:Freiburg10 FACS VP2 A431.png
Media:Freiburg10 FACS VP2Fusion HT1080 Diagramm.png
Media:Freiburg10 Gel VP1ins.png
Media:Freiburg10 GelVP2Fusion2tog.PNG
Media:Freiburg10 GelVP2Fusion2tog2.PNG
Media:Freiburg10 LinkerSchema.png
Media:Freiburg10 NLS mVenus mCherry.png
Media:Freiburg10 NLSSchema.png
Media:Freiburg10 Overview VP1Insertion.png
Media:Freiburg10 Overview VP2Fusion.png
Media:Freiburg10 Primer ColonyPCR.png
Media:Freiburg10 qPCR VP1Insnew.PNG
Media:Freiburg10 qPCR VP2Fus.PNG
Media:Freiburg10 SchemaVP2Fusion.PNG
Media:Freiburg10 TargetingMotifSchema.png
Media:Freiburg10 test supernatant pellet.png
Media:Freiburg10 TestDARPin.png
Media:Freiburg10 TimeLapseA431.png
Media:Freiburg10 TimeLapseHT1080.png
Media:Freiburg10 VP1InsertionSchema.PNG
Media:Freiburg10 VP1upSchema.png
Media:Freiburg10 VP23Schema.png
Media:Freiburg10_N_terminal_fusion_approach.png
Media:Freiburg10_Cotransfected_plasmids.png
VP1 insertion
Media:Freiburg10_pSB1C3_VP1up_NLS_Fig27.png
Media:Freiburg10_VP1_insertion.png
Targeting molecules
Media:Freiburg10_Nucleotide_sequence_ZEGFR.png
Media:Freiburg10_Darpin_internal_repeat_modules.png
Media:Freiburg10_Darpin_conserved_motifs.png
Media:Freiburg10_Darpin_capping_modules.png
Favorite parts
Media:Freiburg10_effect_GCV.png
Media:Freiburg10_viral_brick_587_KO_His.png
Media:Freiburg10_psB1C3_ZEGFR_Linker_VP23.png
Media:Freiburg10_Darpin_internal_repeat_modules.png
Media:Freiburg10_Darpin_conserved_motifs.png
Media:Freiburg10_Darpin_capping_modules.png
HSPG KO
Media:Freiburg10 HSPG binding motif.png
Media:Freiburg10 heparansulfate.PNG
ITR Diary
Targeting: Flow Cytometry: DARPin, Affi (HSPGKO)
Media:Freiburg10 FACS DARPin Affi HSPGKO A431.png
Media:Freiburg10_FACS DARPin Affi HSPGKO HT1080.png
Media:Freiburg10_FlowCytometry Diagramm DARPin Affi.png