User:Nkuldell/mtDNA pt3: Difference between revisions
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(New page: ===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 se...) |
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*Part 1: Rnt1p (RNaseIII enzyme), targetted to mt using sig sequence from HEM1 or COX4. | *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. | *Part 2: Guide RNAs, targetted to mt using lysing-tRNA-CUU (tRK1) import system. | ||
Summary of natural transcriptional control elements [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=2251275&ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Biswas in PNAS 1990 87:9338] and [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=2229061&ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Biswas and Getz in J Biol Chem 1990 265:19053] | |||
===Part 1: Rnt1p=== | ===Part 1: Rnt1p=== | ||
Revision as of 13:39, 13 June 2007
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
Rnt1p
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
- add tag to gene to follow localization of protein product
- microarray induced/uninduced to look for effect of RNase in mt.
- mtBARNase leads to resp- cells when expressed at low level (YPEG from GALS promoter) and is toxic at high levels (SC-U/Gal), unless simultanously express mtBARSTAR inhibitor protein Mireau, Arnal and Fox in Mol Gen Genomics 2003 270:1-8.
