User:Nkuldell/mtDNA pt3

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===Overall goal===
===Overall goal===
Make a tool to regulate gene expression of any mt gene. Tool would have two-parts and be nuclear-encoded.  
Make a tool to regulate gene expression of any mt gene. Tool would have two-parts and be nuclear-encoded.  
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*Part 1: Rnt1p (RNaseIII enzyme), targetted to mt using sig sequence from HEM1 or COX4.
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*Part 1: [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=rnt1 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.
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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]
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Foundational info: 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]<br>
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Foundational info: works for degradation of mRNAs in S. cerevisiae nucleus [http://www.plosone.org/article/fetchArticle.action;jsessionid=52A54F60AD8F8DBEB598BBCBDBD16464?articleURI=info%3Adoi%2F10.1371%2Fjournal.pone.0000472 Lamontagne and Elela in PLoS One 2007 5:e472]
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===Part 1: Rnt1p===
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===Part 1: [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=rnt1 Rnt1p]===
====Gen'l info about RNases====
====Gen'l info about RNases====
====S. cerevisiae RNAases====
====S. cerevisiae RNAases====

Revision as of 16:47, 13 June 2007

Contents

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.

Foundational info: Summary of natural transcriptional control elements Biswas in PNAS 1990 87:9338 and Biswas and Getz in J Biol Chem 1990 265:19053
Foundational info: works for degradation of mRNAs in S. cerevisiae nucleus Lamontagne and Elela in PLoS One 2007 5:e472

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

    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

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