User:Nkuldell/mtDNA pt3

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Overall goal

Make a tool to regulate gene expression of any mt gene. Tool would have two-parts and be nuclear-encoded.

  • Part 1: Rnt1p (RNaseIII enzyme), targetted to mt using sig sequence from HEM1 or COX4.
  • Part 2: Guide RNAs, targetted to mt using lysing-tRNA-CUU (tRK1) import system.

Summary of natural transcriptional control elements Biswas in PNAS 1990 87:9338 and Biswas and Getz in J Biol Chem 1990 265:19053

Part 1: Rnt1p

Gen'l info about RNases

S. cerevisiae RNAases


Part 2: Guide RNAs

Structural requirements for guide RNAs

Cell components needed for moving guide RNA to mt

  • piggy back

Experimental checkpoints for regulated expression

Part 1: Rnt1p in mt

    1. add tag to gene to follow localization of protein product
    2. microarray induced/uninduced to look for effect of RNase in mt.

Applications for mt gene regulation, in the unlikely case that this system works