Featured Parts:RNA-Lock-and-Key

Revision as of 06:15, 27 April 2006 by Smelissali (Talk | contribs) (→‎Background: How the Riboregulator Works)

The Parts

Bba_J01008 - ("key 1") Biobricked version of Isaacs' riboregulator trans activating key, taR12
Bba_J01010 - ("lock 1") Biobricked version of Isaacs' riboregulator cis repressed lock, crR12.
Bba_J01080 - ("lock 3") Modified version of J01010
Bba_J01086 - ("key 3") Modified version of J01008

Background: How the Riboregulator Works

This set of riboregulators are based upon the work of Isaacs et al 2004 (see reference #1). They work by repressing the expression of a target gene by introducing a complementary region to prevent transcription.
The DNA sequences of the "locks" are modified directly upsteam of the target gene's ribosome binding site (RBS) such that the 5' untranslated region will fold to form a stem-loop secondary structure, thus blocking the RBS and preventing transcription. This stem-loop structure is known as the 'cis-repressed' mRNA (crRNA).
These "locked" stem-loops are 'unlocked' via what we call "keys". These "keys" f non-coding RNA are called "trans-activating" RNA (taRNA). They act to bind complementary regions within the lock to open the crRNA and allow proteins to bind at the RBS.

Suggested Uses for these Biobricks

Experimental and Technical

Technical

Previous Experiments

References

  • Publications on the Isaacs Riboregulator:
    • Isaacs, Dwyer DJ, Ding C, Pervouchine DD, Cantor CR, Collins JJ. 2004. "Engineered riboregulators enable post-transcriptional control of gene expression". Nature Biotechnology 2004. [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15208640&query_hl=7&itool=pubmed_docsum Pubmed] [http://www.nature.com/nbt/journal/v22/n7/abs/nbt986.html;jsessionid=ABB92F041E994E3B8E4A5213699C3CB0 Journal]
  • More parts and resources
    • A listing of more parts made by the Berkeley 2005 iGEM team including constituitively 'on' promoter-controlled riboregulators and other experimental constructs can be found here