User:Pakpoom Subsoontorn/Notebook/Genetically Encoded Memory/2008/10/11

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Expanding recombinase library

  • For this project, we will evolve a large library of recombinase system whose target sequences do not exist in nature. The experiment will involve protein evolution, high-throughput screening assay and bioinformatics. We also want to learn what set of sequence cannot be used as a target.
  • Goal: higher efficiency, irreversibility, change of site-specificity

Project outline

Experiment setup

Questions

  • What're the general limitations of protein evolution? What could be the particular limitations in evolving recombinases? Library site? Screening?
  • Not all sequences are physically capable for being recombinase sites. How can we explore many different evolution paths simultaneously? Let's say we have 100,000 mutant in the library. Maybe there are hundreds of promising new sequences that are not natural recombination target sequences.

TO DO list

Reference