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<img align="center" src="https://static.igem.org/mediawiki/2016/1/11/DcasPlasmidTrans.png" alt = "dCas9Plasmid" /> | <img align="center" src="https://static.igem.org/mediawiki/2016/1/11/DcasPlasmidTrans.png" alt = "dCas9Plasmid" /> | ||
− | <h4 align="center"> This is our first plasmid. It contains one of the editing enzymes, either ADAR1, ADAR2, or APOBEC1, fused to dCas9. Downstream on the same plasmid is the rtTA gene and promoter. </h4> | + | <h4 align="center"> This is our first plasmid. It contains one of the editing enzymes, either ADAR1, ADAR2, or APOBEC1, fused to dCas9. The enzyme and dCas9 have 1-3 repeats of an XTEN linker to allow the enzyme to edit a certain distance away from dCas9. Downstream on the same plasmid is the rtTA gene and promoter. </h4> |
<img align="center" src="https://static.igem.org/mediawiki/2016/6/69/TunabilityTrans.png" alt = "tunability" /> | <img align="center" src="https://static.igem.org/mediawiki/2016/6/69/TunabilityTrans.png" alt = "tunability" /> | ||
− | <h4 align="center"> One of the key parts of our system is that we wanted it to be tunable. We don't want the enzymes to be editing at all time, especially if something was to go wrong. Regulation is the purpose of the rtTA gene in this plasmid. rtTA, which stands for reverse tetracycline trans activator, is constitutively expressed. It acts as a repressor binding to the tetracycline responsive element (TRE) promoter and preventing transcription. However, when doxycycline is added to the system it will bind to the rtTA, removing it from the promoter and allowing transcription. Editing will not occur unless we introduce doxycycline into the cells. </h4> | + | <h4 align="center"> One of the key parts of our system is that we wanted it to be tunable. We don't want the enzymes to be editing at all time, especially if something was to go wrong. Regulation is the purpose of the rtTA gene in this plasmid. rtTA, which stands for reverse tetracycline trans activator, is constitutively expressed. It acts as a repressor binding to the tetracycline responsive element (TRE) promoter and preventing transcription of the editing enzyme and dCas9. However, when doxycycline is added to the system it will bind to the rtTA, removing it from the promoter and allowing transcription. Editing will not occur unless we introduce doxycycline into the cells. </h4> |
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Revision as of 20:53, 14 October 2016