IGEM:Caltech/2007/Project: Difference between revisions
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As an initial mechanism to target viruses to specific cell types, we will place the viral developmental genes under riboregulator control. Viral mRNAs for the regulated developmental genes will express with a stem loop sequestering ribosome binding sites, preventing translation. Specific mRNA in target E. coli will invade the stem loop, freeing the ribosome binding site and allowing proper translation. We believe this approach is more general than methods which might target specific cell-surface markers. Furthermore, if this method works, it would be possible in principle to extend viral mRNA regulation using aptamers capable of recognizing subtle signals such as post-translational modification. | As an initial mechanism to target viruses to specific cell types, we will place the viral developmental genes under riboregulator control. Viral mRNAs for the regulated developmental genes will express with a stem loop sequestering ribosome binding sites, preventing translation. Specific mRNA in target E. coli will invade the stem loop, freeing the ribosome binding site and allowing proper translation. We believe this approach is more general than methods which might target specific cell-surface markers. Furthermore, if this method works, it would be possible in principle to extend viral mRNA regulation using aptamers capable of recognizing subtle signals such as post-translational modification. | ||
We selected the viral developmental genes <i>N, Q,</i> and <cro> as promising targets for regulation. <i>N</i> and <i>Q</i> are <i>antiterminators</i> required for λ to transcribe its full set of genes. Viruses lacking these genes stall at extremely early developmental stages and are completely inviable, barely producing any viral mRNA. <i>cro</i> represents a potentially interesting means to bias whether the virus will lyse a target cell, or integrate into its DNA. This makes it an attractive candidate to investigate the rewiring applications mentioned above. | We selected the viral developmental genes <i>N, Q,</i> and <i>cro</i> as promising targets for regulation. <i>N</i> and <i>Q</i> are <i>antiterminators</i> required for λ to transcribe its full set of genes. Viruses lacking these genes stall at extremely early developmental stages and are completely inviable, barely producing any viral mRNA. <i>cro</i> represents a potentially interesting means to bias whether the virus will lyse a target cell, or integrate into its DNA. This makes it an attractive candidate to investigate the rewiring applications mentioned above. | ||
Choosing an appropriate λ strain poses a challenge. Existing strains with defective <i>N, Q,</i> and <i>cro</i> genes lack unique restriction sites to clone our constructs into. Therefore, we will utilize recombineering to introduce introduce these mutations into phages specifically designed to accept cloning inserts. | |||
Working with lambda phage has been new and fun for all of us | Working with lambda phage has been new and fun for all of us. We hope you take the time to browse our wiki and learn more about our work! | ||
[[Image:Caltech_2007_overview.gif|center]] | [[Image:Caltech_2007_overview.gif|center]] |
Revision as of 00:22, 26 October 2007
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