Team:Freiburg Bioware/BioBricks/capsid coding


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Latest revision as of 01:05, 28 October 2010

Capsid Coding

BBa_K404001[AAV2]-Rep-VP123Team Freiburg 20104214
BBa_K404002[AAV2]-Rep-VP123_p5-TATAlessFreiburg Bioware 20104386
BBa_K404003[AAV2]-Rep-VP123(ViralBrick-587KO-empty)_p5-TATAlessFreiburg Bioware 20104386
BBa_K404004[AAV2]-Rep-VP13(ViralBrick-587KO-empty)_p5-TATAlessFreiburg Bioware 20104402
BBa_K404005[AAV2]-Rep-VP23(ViralBrick-587KO-empty)_p5-TATAlessFreiburg Bioware 20104402
BBa_K404006[AAV2]-VP123Freiburg Bioware 20102208
BBa_K404007pCMV_[AAV2]-VP123Freiburg Bioware 20102998
BBa_K404008[AAV2]-VP1Freiburg Bioware 20102202
BBa_K404009pCMV_[AAV2]-VP1Freiburg Bioware 20102910
BBa_K404010[AAV2]-VP2Freiburg Bioware 20101830
BBa_K404011pCMV_[AAV2]-VP2Freiburg Bioware 20102500
BBa_K404012[AAV2]-VP3Freiburg Bioware 20101635
BBa_K404013pCMV_[AAV2]-VP3Freiburg Bioware 20102305
BBa_K404150[AAV2]-VP23 Freiburg Bioware 20101917
BBa_K404151[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20101917
BBa_K404152[AAV2]-VP1up Freiburg Bioware 2010411
BBa_K404153[AAV2]-NLS Freiburg Bioware 201021
BBa_K404154pCMV_Z-EGFR-1907_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102767
BBa_K404155pCMV_Z-EGFR-1907_Short-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102785
BBa_K404156pCMV_Z-EGFR-1907_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102797
BBa_K404157pCMV_Z-EGFR-1907_Long-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102809
BBa_K404158pCMV_Z-EGFR-1907_SEG-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102881
BBa_K404159pCMV_CFP_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20103337
BBa_K404160pCMV_His-Tag_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20102641
BBa_K404161pCMV_DARPin-E01_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20103082
BBa_K404162pCMV_Z-EGFR-1907_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-BAP) Freiburg Bioware 20102857
BBa_K404163pCMV_Z-EGFR-1907_Middle-Linker_[AAV2]-VP23 (ViralBrick-587KO-His-Tag) Freiburg Bioware 20102830
BBa_K404164pCMV_VP1up_NLS_Z-EGFR-1907_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20103211
BBa_K404165pCMV_VP1up_NLS_mVenus_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20103751
BBa_K404166pCMV_VP1up_NLS_His-Tag_[AAV2]-VP23 (ViralBrick-587KO-Empty) Freiburg Bioware 20103056
BBa_K404167pCMV_VP1up_NLS_mVenus_VP23(ViralBrick-587-KO-BAP)Freiburg Bioware 20103811
BBa_K404168pCMV_VP1up_NLS_mVenus_VP2/3(ViralBrick-587-KO-His)Freiburg Bioware 20103784
BBa_K404204ViralBrick-453-Z34C Freiburg Bioware 2010165
BBa_K404206ViralBrick-587-His-Tag Freiburg Bioware 2010105
BBa_K404210ViralBrick-587KO-Empty Freiburg Bioware 201072
BBa_K404222[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-453-His-Tag) Freiburg Bioware 20104410
BBa_K404223[AAV2]-Rep-VP123_p5-TATAless (ViralBrick-453-RGD) Freiburg Bioware 20104419
BBa_K404224[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-453-Z34C) Freiburg Bioware 20104488
BBa_K404225[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-587-BAP) Freiburg Bioware 20104446
BBa_K404227[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-587-RGD) Freiburg Bioware 20104428
BBa_K404229[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-587KO-BAP) Freiburg Bioware 20104446
BBa_K404232[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-587KO-RGD) Freiburg Bioware 20104428
BBa_K404234[AAV2]-Rep-VP123_P5-TATAless (ViralBrick-587KO-Z34C-Spacer) Freiburg Bioware 20104503
BBa_K404235[AAV2]Rep-VP123_p5TATAless (ViralBrick 453 Z34C-587 HSPG-KO)Freiburg Bioware 20104488
BBa_K404236[AAV2]Rep-VP123_p5TATAless (ViralBrick 453 RGD-587 HSPG-KO)Freiburg Bioware 20104419
BBa_K404241pCMV_[AAV2]VP123 (ViralBrick-453-BAP) Freiburg Bioware 20103049
BBa_K404242pCMV_[AAV2]VP123 (ViralBrick-453-His-Tag)Freiburg Bioware 20103022
BBa_K404243pCMV_[AAV2]VP123 (ViralBrick-453-RGD)Freiburg Bioware 20103031
BBa_K404244pCMV_[AAV2]VP123 (ViralBrick-453-Z34C) Freiburg Bioware 20103100
BBa_K404246pCMV_[AAV2]-VP123ex (ViralBrick-587-BAP)Freiburg Bioware 20103058
BBa_K404247pCMV_[AAV2]-VP123ex (ViralBrick-587-His-Tag)Freiburg Bioware 20103031
BBa_K404248pCMV_[AAV2]-VP123ex (ViralBrick-587-RGD)Freiburg Bioware 20103040
BBa_K404250pCMV_[AAV2]-VP123ex (ViralBrick-587-Beta-lactamase)Freiburg Bioware 20103787
BBa_K404251pCMV_[AAV2]-VP123ex (ViralBrick-587KO-BAP)Freiburg Bioware 20103058
BBa_K404252pCMV_[AAV2]-VP123ex (ViralBrick-587KO-Empty)Freiburg Bioware 20102998
BBa_K404253pCMV_[AAV2]-VP123ex (ViralBrick-587KO-His-Tag)Freiburg Bioware 20103031
BBa_K404254pCMV_[AAV2]-VP123ex (ViralBrick-587KO-RGD)Freiburg Bioware 20103040
BBa_K404255pCMV_[AAV2]-VP123ex (ViralBrick-587KO-Z34C)Freiburg Bioware 20103100
BBa_K404256pCMV_[AAV2]-VP123ex (ViralBrick-587KO-Z34C-Spacer)Freiburg Bioware 20103115
BBa_K404314DARPin-E01 Freiburg Bioware 2010459

Basic Capsidcoding Constructs

The category of basic capsid coding constructs contains different BioBricks that can be used to produce different kinds of viral therapy vectors. All of these constructs are compatible to the ViralBrick standard and can therefor be modifyed in a single cloning step to contain functional peptide in the major surface exposed loops.

The Basic Capsid Coding Parts

  • [AAV-2]-Rep-VP123_p5-TATA-less is the main capsid coding plasmid.
  • pCMV_[AAV-2]-VP123 is ment for co-transfections aiming to produce mosaic viruses.
  • pCMV_[AAV-2]-VP13 and pCMV_[AAV-2]-VP13 are ment for co-transfections when using N-terminal fusion.
  • pCMV_[AAV-2]-VP1, pCMV_[AAV-2]-VP2 and pCMV_[AAV-2]-VP3 are constructs code for only one of the three viral coat proteins and can be used to produce the viral coat portein, to purify them and to perform InVitro assembly of the viral shell.

ViralBricks for Loop replacement

ViralBricks are short nucleotide sequences derived from the gene of the viral coat proteins VP1-3 where they encode the two major surface exposed loops around the positions 453 and 587. Functional peptides are integrated in the loop tip which will be present on the surface of viral vectros that have been modified with the corresponding ViralBrick.

Different ViralBrick versions:

Different functional motifs:

Versions of the Rep-Vp123_p5-TATA-less

The construct [AAV-2]-RepVP123_p5-TATA-less is the construct that is cloned from an AAV2 Helper-free System which is derived from the genome of the Adeno-associated Virus in which the ITRs were removed. The expression of the eight gene products is regulated by three promoters. The p40 regulates the expression of the viral coat proteins and the AAP whereas the promoters p5 and p19 regulate the expression of the Rep proteins that are important for the production of the vector plasmid and the packaging. The usage of a TATA-less p5 promoter that is located downstream of the open reading frames favoures the production of the short Rep variants over the long ones which is conductive for the production of viral vectors.

Versions of the pCMV_Vp123

The different versions of pCMV_VP123 are ment for the production of mosaic viruses by transfecting this plamid together with [AAV2]-Rep-VP123_P5-TATA-less. The composite part pCMV_VP123 contains the ORF of the Viral Coat Proteins VP123 under the control of the control of the Cytomegalovirus promoter. By providing modified and unmodified viral coat proteins the resulting viral vector will be a chimera. The ratio of modified to unmodified viral coat proteins will depend on the number of plasmids a cell was transfected with and has impect on the functionality of the viral vector.