Team:Freiburg Bioware/Filelist

From 2010.igem.org

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).

Contents

1 AAAAAH

2 eGFP

3 Time lapse

4 Structure Modeling

5 Modularaization of GOI - Results

6 Introduction to Adeno-Associated Virus Serotype 2

7 Trafficking

8 Helper genes

9 Gene therapy

10 Cytosine deaminase

11 Loop insertions

12 Tagging

13 Targeting

14 N-terminal fusion

14.1 Results

15 VP1 insertion

16 Targeting molecules

17 Favorite parts

18 HSPG KO

19 ITR Diary

20 Targeting: Flow Cytometry: DARPin, Affi (HSPGKO)

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

Media:Freiburg10_Rep40.jpg

Trafficking

Media:Freiburg10_viral_trafficking.png

Media:Freiburg10_cellular_surface_HSPG.png

Media:Freiburg10_Clathrin_triskelions.png

Helper genes

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Media:Freiburg10_pHelper.png

Gene therapy

Media:Freiburg10_overview_GDEPT.png

Media:Freiburg10_Bystander_effect.png

Cytosine deaminase

Media:Freiburg10_Flourocytosine_pathway.png

Media:Freiburg10_CD.jpg

Media:Freiburg10_TK.jpg

Loop insertions

Media:Freiburg10_3D_AC_of_453_587_loops.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

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Media:Freiburg10_BAP-tag.png

Media:Freiburg10_BAP-experiment.png

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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

Tagging

Media:Freiburg10_BAP.png

Media:Freiburg10_His.png

Targeting

Media:Freiburg10_RGD_motif.png

Media:Freiburg10_Z34C_motif.png

Media:Freiburg10_Z34C_Z38_ZDomain_helix.png

Media:Freiburg10_HSPG.png

Media:Freiburg10_HSPG_chemical_structure.png

N-terminal fusion

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_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.png

Media:Freiburg10_Darpin_internal_repeat_modules.png

Media:Freiburg10_Darpin_conserved_motifs.png

Media:Freiburg10_Darpin_capping_modules.png

Favorite parts

Media:Freiburg10_IMAC.png

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

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