Difference between revisions of "Part:BBa K1442006"
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<p>An aptazyme is an allosteric ribozyme acting as a regulatory riboswitch. It self-splices in response to theophylline, a drug used to treat asthma and other chronic obstructive pulmonary diseases acting as a bronchodilator. This differs from caffeine by one methyl group and a nitrogen atom creating the structure seen below. This is not endogenous to the body therefore we used this element as a "kill switch" for our system.</p> | <p>An aptazyme is an allosteric ribozyme acting as a regulatory riboswitch. It self-splices in response to theophylline, a drug used to treat asthma and other chronic obstructive pulmonary diseases acting as a bronchodilator. This differs from caffeine by one methyl group and a nitrogen atom creating the structure seen below. This is not endogenous to the body therefore we used this element as a "kill switch" for our system.</p> | ||
| − | <p> | + | <p><html><body><img src="https://static.igem.org/mediawiki/parts/5/5d/Aptazyme.jpg"/></body></html></p> |
| − | + | <p><i><html><body><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC139998/#!po=13.2353">Group I aptazymes as gentic regulatory switches</a></body></html> The aptazyme we used was from the Group I intron-splicing type. 5' and 3' splice sites are indicated with arrows and indicated where the ribozyme self-splices following binding of a theophylline molecule.</i></p> | |
| − | <!-- Add more about the biology of this part here | + | |
| − | + | <p>One RNA molecule binds to theophylline with a dissociation constant of 0.1microM and a significant conformational change occurs on binding. The addition of the aptamer to the ribozyme significantly increases the activation of the ribozyme when theophylline is added, creating a ribozyme activated 1600 times in saturating theophylline concentrations. [<html><body><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC139998/#!po=13.2353">THompson et al. 2002</a></body></html>] When included in a system they can be used as knockouts for certain mRNAs or to make more economical gene therapies.</p> | |
| + | <!-- Add more about the biology of this part here--> | ||
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| + | == Usage == | ||
| + | <p>This aptazyme was included as an RNA kill switch We put this structure centrally in our replicon in order to cause the maxmimum disruption when activated with theophylline.</p> | ||
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Revision as of 20:55, 11 October 2014
Anti-theophylline Aptazyme
An aptazyme is an allosteric ribozyme acting as a regulatory riboswitch. It self-splices in response to theophylline, a drug used to treat asthma and other chronic obstructive pulmonary diseases acting as a bronchodilator. This differs from caffeine by one methyl group and a nitrogen atom creating the structure seen below. This is not endogenous to the body therefore we used this element as a "kill switch" for our system.
Group I aptazymes as gentic regulatory switches The aptazyme we used was from the Group I intron-splicing type. 5' and 3' splice sites are indicated with arrows and indicated where the ribozyme self-splices following binding of a theophylline molecule.
One RNA molecule binds to theophylline with a dissociation constant of 0.1microM and a significant conformational change occurs on binding. The addition of the aptamer to the ribozyme significantly increases the activation of the ribozyme when theophylline is added, creating a ribozyme activated 1600 times in saturating theophylline concentrations. [
THompson et al. 2002] When included in a system they can be used as knockouts for certain mRNAs or to make more economical gene therapies.Usage
This aptazyme was included as an RNA kill switch We put this structure centrally in our replicon in order to cause the maxmimum disruption when activated with theophylline.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]