Difference between revisions of "Part:BBa K404203"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K404203 short</partinfo>
 
<partinfo>BBa_K404203 short</partinfo>
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="left"
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! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404203 ViralBrick-453-RGD]
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|-
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|'''BioBrick Nr.'''
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|[https://parts.igem.org/Part:BBa_K404203 BBa_K404203]
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
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|-
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|'''Requirement'''
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|pSB1C3<br>
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|-
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|'''Source'''
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|
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|-
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|'''Submitted by'''
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|[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]
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|}
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<br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/>
  
 
<b>The RGD integrin binding motif, ready for insertion into the 453 loop</b>
 
<b>The RGD integrin binding motif, ready for insertion into the 453 loop</b>
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https://static.igem.org/mediawiki/parts/f/f6/Freiburg10_ViralBrick_motif_RGD.png
 
https://static.igem.org/mediawiki/parts/f/f6/Freiburg10_ViralBrick_motif_RGD.png
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 +
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===Restriction sites===
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<table border="1" style="float:right; width:480px; height:auto;>
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<tr>
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  <th><b>Restriction site</b></th>
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  <th><b>Enzyme</b></th>
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  <th><b>Recognition sequence</b></th>
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  <th><b>Activity</b></th>
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  <th><b>Temp.</b></th>
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</tr>
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<tr>
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  <td>upstream 453</td>
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  <td>[http://www.neb.com/nebecomm/products/productR3132.asp SspI-HF]</td>
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  <td>[[image:Freiburg10_recognition site SspI-HF.Gif]]</td>
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  <td>25;100;0;100</td>
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<td>37°C</td>
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</tr>
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<tr>
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  <td>downstream 453</td>
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  <td>[http://www.neb.com/nebecomm/products/productR3138.asp SalI-HF]</td>
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  <td>[[image:Freiburg10_recognition site SalI-HF.Gif]]</td>
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  <td>10;100;100;100</td>
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  <td>37°C</td>
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<tr>
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</table>
 
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Latest revision as of 02:20, 28 October 2010

ViralBrick-453-RGD

ViralBrick-453-RGD
BioBrick Nr. BBa_K404203
RFC standard RFC 25
Requirement pSB1C3
Source
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]














The RGD integrin binding motif, ready for insertion into the 453 loop

Freiburg10 ViralBrick-logo-453-RGD.png


Usage and Biology

Integrins are transmembrane proteins that, among other functions, mediate cell attachment to surrounding tissues. They bind to a motif consisting of the amino acids arginine, glycine and aspartic acid (RGD in one-letter code). Because Integrin is highly expressed in many tumor cell lines (Albelda et al., 1990), (Damjanovich, Albelda, Mette, & Buck, 1992), (Lessey et al., 1995), (Smythe, LeBel, Bavaria, Kaiser, & Albelda, 1995), (Gladson & Cheresh, 1991), AAV particles displaying the RGD motif on various positions in their capsid proteins have been created by (Shi et al., 2003). Particles displaying RGD at amino acid positions 584 & 588 as well as 453 or 587 (Boucas et al., 2009) showed transduction efficiencies similar to wt AAV, even when the cells’ HSPG receptors were blocked by heparin sulfate or when the natural HSPG binding motif on the capsid surface was knocked out. To further broaden the area of therapeutic application, we created a ViralBrick containing the RGD motive to specifically target cells with low HSPG-/high Integrin expression.

Freiburg10_ViralBrick_motif_RGD.png


Restriction sites

Restriction site Enzyme Recognition sequence Activity Temp.
upstream 453 [http://www.neb.com/nebecomm/products/productR3132.asp SspI-HF] Freiburg10 recognition site SspI-HF.Gif 25;100;0;100 37°C
downstream 453 [http://www.neb.com/nebecomm/products/productR3138.asp SalI-HF] Freiburg10 recognition site SalI-HF.Gif 10;100;100;100 37°C

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]