Ljubljana/summary

From 2007.igem.org

(Difference between revisions)
m
Line 36: Line 36:
   <li>Design and implementation of three functional systems of activation of anti-HIV defense based on:
   <li>Design and implementation of three functional systems of activation of anti-HIV defense based on:
   <dl>
   <dl>
-
     <dd>- the detection of viral interactions with the cells and
+
     <dd>- detection of viral interactions with the cells and
     <dd>- cleavage of specific substrate by viral protease.
     <dd>- cleavage of specific substrate by viral protease.
   </dl>
   </dl>
Line 44: Line 44:
     <dd>- split ubiquitin system.
     <dd>- split ubiquitin system.
   </dl>
   </dl>
-
   <li>Construction of <a href="https://2007.igem.org/Ljubljana/biobricks">BioBricks</a> for heterodimerization, which represent a powerful platform for engineering cell processes based on protein-protein interactions.
+
   <li>Construction of <a href="https://2007.igem.org/Ljubljana/biobricks">BioBricks</a> for heterodimerization, which represents a powerful platform for engineering cell processes based on protein-protein interactions.
-
   <li>Introduction of the versatile amplification platform based on the use of T7 RNA polymerase in mammalian cells.
+
   <li>Introduction of a versatile amplification platform based on the use of T7 RNA polymerase in mammalian cells.
-
   <li>Activation device based on the proteolytic cleavage and release of membrane anchored T7 RNA polymerase from the membrane into the cell nucleus.
+
   <li>Activation device based on the proteolytic cleavage and release of membrane-anchored T7 RNA polymerase from the membrane into the cell nucleus.
   <li>Anchoring of T7 RNA polymerase to the plasma membrane either by:
   <li>Anchoring of T7 RNA polymerase to the plasma membrane either by:
   <dl>
   <dl>
Line 54: Line 54:
   <li>Implementation of two different types of effector molecules for:  
   <li>Implementation of two different types of effector molecules for:  
   <dl>
   <dl>
-
     <dd>- induction of cell death (caspase-3)or
+
     <dd>- induction of cell death (caspase-3) or
     <dd>- activation of antiviral defense (interferon beta).
     <dd>- activation of antiviral defense (interferon beta).
   </dl>
   </dl>
Line 77: Line 77:
<li>The idea of relying on viral functions could be applied also to other HIV-specific processes (such as genome integration, reverse transcription…).
<li>The idea of relying on viral functions could be applied also to other HIV-specific processes (such as genome integration, reverse transcription…).
<li>Our system based on <a href="https://2007.igem.org/Ljubljana/biobricks">BioBricks</a> has been useful to prove the principle, but for the real application cellular delivery would be much better applied using vectors based on viral delivery.
<li>Our system based on <a href="https://2007.igem.org/Ljubljana/biobricks">BioBricks</a> has been useful to prove the principle, but for the real application cellular delivery would be much better applied using vectors based on viral delivery.
-
<li>In the case of using viral vectors, therapy could be delivered specifically to the relevant cells that can be infected by HIV-1 (e.g. similar as T-cell-directed retroviral delivery systems described by David Baltimore at SB2.0).
+
<li>In the case of using viral vectors, therapy could be targeted specifically to cells that can be infected by HIV-1 (e.g. similar to T-cell-directed retroviral delivery systems described by David Baltimore at SB2.0).
<li>Selection of different effectors will (and can) be made based on further experiments – apoptosis can only be used in 100% leak-proof systems, while antiviral and immune response effectors are not as sensitive.
<li>Selection of different effectors will (and can) be made based on further experiments – apoptosis can only be used in 100% leak-proof systems, while antiviral and immune response effectors are not as sensitive.
<li>Similar systems can be prepared for therapy of many other pathogens, particularly viruses (e.g. yellow fever, SARS, foot and mouth disease…) which contain specific proteases and rely on defined cellular receptors.
<li>Similar systems can be prepared for therapy of many other pathogens, particularly viruses (e.g. yellow fever, SARS, foot and mouth disease…) which contain specific proteases and rely on defined cellular receptors.

Revision as of 18:03, 26 October 2007

Company Name

Achievements

  • Design and implementation of three functional systems of activation of anti-HIV defense based on:
    - detection of viral interactions with the cells and
    - cleavage of specific substrate by viral protease.
  • Successful detection of CD4-CCR5 heterodimer formation through:
    - split tobacco etch virus (TEV) protease,
    - split ubiquitin system.
  • Construction of BioBricks for heterodimerization, which represents a powerful platform for engineering cell processes based on protein-protein interactions.
  • Introduction of a versatile amplification platform based on the use of T7 RNA polymerase in mammalian cells.
  • Activation device based on the proteolytic cleavage and release of membrane-anchored T7 RNA polymerase from the membrane into the cell nucleus.
  • Anchoring of T7 RNA polymerase to the plasma membrane either by:
    - fusion to CD4 or
    - addition of a myristoylation signal.
  • Implementation of two different types of effector molecules for:
    - induction of cell death (caspase-3) or
    - activation of antiviral defense (interferon beta).

Prospects for future work and real world applications

  • The idea of relying on viral functions could be applied also to other HIV-specific processes (such as genome integration, reverse transcription…).
  • Our system based on BioBricks has been useful to prove the principle, but for the real application cellular delivery would be much better applied using vectors based on viral delivery.
  • In the case of using viral vectors, therapy could be targeted specifically to cells that can be infected by HIV-1 (e.g. similar to T-cell-directed retroviral delivery systems described by David Baltimore at SB2.0).
  • Selection of different effectors will (and can) be made based on further experiments – apoptosis can only be used in 100% leak-proof systems, while antiviral and immune response effectors are not as sensitive.
  • Similar systems can be prepared for therapy of many other pathogens, particularly viruses (e.g. yellow fever, SARS, foot and mouth disease…) which contain specific proteases and rely on defined cellular receptors.