Difference between revisions of "Part:BBa K404203"

Latest revision as of 02:20, 28 October 2010

ViralBrick-453-RGD

The RGD integrin binding motif, ready for insertion into the 453 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 453	[http://www.neb.com/nebecomm/products/productR3132.asp SspI-HF]		25;100;0;100	37°C
downstream 453	[http://www.neb.com/nebecomm/products/productR3138.asp SalI-HF]		10;100;100;100	37°C

Sequence and Features