Difference between revisions of "Part:BBa K404207"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K404207 short</partinfo> | <partinfo>BBa_K404207 short</partinfo> | ||
| + | {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="left" | ||
| + | ! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404207 ViralBrick-587-RGD] | ||
| + | |- | ||
| + | |'''BioBrick Nr.''' | ||
| + | |[https://parts.igem.org/Part:BBa_K404207 BBa_K404207] | ||
| + | |- | ||
| + | |'''RFC standard''' | ||
| + | |[https://parts.igem.org/Help:Assembly_standard_25 RFC 25] | ||
| + | |- | ||
| + | |'''Requirement''' | ||
| + | |pSB1C3<br> | ||
| + | |- | ||
| + | |'''Source''' | ||
| + | | | ||
| + | |- | ||
| + | |'''Submitted by''' | ||
| + | |[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010] | ||
| + | |} | ||
| + | <br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/> | ||
<br><b>The RGD integrin binding motif, ready for insertion into the 587 loop</b> | <br><b>The RGD integrin binding motif, ready for insertion into the 587 loop</b> | ||
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<div style="float:left; width:460px; height:auto; margin: 0px 5px 0px 5px; text-align:justify;"> | <div style="float:left; width:460px; height:auto; margin: 0px 5px 0px 5px; text-align:justify;"> | ||
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. | 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. | ||
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</div> | </div> | ||
<div style="float:right; width:480px; height:auto; "><img src="https://static.igem.org/mediawiki/parts/f/f6/Freiburg10_ViralBrick_motif_RGD.png" width="460" | <div style="float:right; width:480px; height:auto; "><img src="https://static.igem.org/mediawiki/parts/f/f6/Freiburg10_ViralBrick_motif_RGD.png" width="460" | ||
height="auto"/></div></div> | height="auto"/></div></div> | ||
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</html> | </html> | ||
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<tr> | <tr> | ||
</table> | </table> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K404207 SequenceAndFeatures</partinfo> | <partinfo>BBa_K404207 SequenceAndFeatures</partinfo> | ||
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<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K404207 parameters</partinfo> | <partinfo>BBa_K404207 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
Latest revision as of 02:29, 28 October 2010
ViralBrick-587-RGD
| ViralBrick-587-RGD | |
|---|---|
| BioBrick Nr. | BBa_K404207 |
| 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 587 loop
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.
Restriction sites
| Restriction site | Enzyme | Recognition sequence | Activity | Temp. |
|---|---|---|---|---|
| upstream 587 | [http://www.neb.com/nebecomm/products/productR3136.asp BamHI-HF] |  |
100;50;10;100 | 37°C |
| downstream 587 | [http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF] |  |
0;25;0;100 | 37°C |
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 10
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]