Difference between revisions of "Part:BBa K3017070"

Revision as of 05:50, 18 October 2019

Combined CRISPRi and antisense RNA Toggle Switch - Model circuit

This construct is a proof of concept of a biological switch, novel to iGEM community, by our team. Our team has combined the CRISPRi system with RNA regulators to achieve a toggle switch. The switch utilizes the catalytically inactive form of Cas9 (dCas9) to achieve targeted and reversible repression of genes via specific single-guide RNAs (sgRNAs). Alternatively, the transcription of antisense RNA (asRNAs) reverses the effect of the dCas9 modulated repression on the desired genes. This method of regulation would allow for the ability to fine-tune and easily customize the execution of highly complex genetic circuits.

To bring our project to life, we have designed a circuit that drives the Escherichia coli (E.coli) strain DH5-alpha to alternately produce green fluorescent protein (GFP) and red fluorescent protein (RFP) depending on the inducer added to our system.

Proposed Toggle Switch Construct

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 852
Illegal PstI site found at 2274
Illegal PstI site found at 2478
Illegal PstI site found at 2508
Illegal PstI site found at 3720
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 5543
Illegal NheI site found at 6921
Illegal NheI site found at 7093
Illegal NheI site found at 7116
Illegal NheI site found at 8733
Illegal PstI site found at 852
Illegal PstI site found at 2274
Illegal PstI site found at 2478
Illegal PstI site found at 2508
Illegal PstI site found at 3720
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 313
Illegal BamHI site found at 5482
Illegal BamHI site found at 6860
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 852
Illegal PstI site found at 2274
Illegal PstI site found at 2478
Illegal PstI site found at 2508
Illegal PstI site found at 3720
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 852
Illegal PstI site found at 2274
Illegal PstI site found at 2478
Illegal PstI site found at 2508
Illegal PstI site found at 3720
Illegal NgoMIV site found at 1140
Illegal NgoMIV site found at 2244
Illegal NgoMIV site found at 2317
Illegal NgoMIV site found at 2802
Illegal NgoMIV site found at 3711
Illegal AgeI site found at 5317
Illegal AgeI site found at 6695
Illegal AgeI site found at 7702
Illegal AgeI site found at 7814
Illegal AgeI site found at 9342
Illegal AgeI site found at 9454
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 5299
Illegal SapI site found at 6677

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3017070 short</partinfo>
-/
+<p> This construct is a proof of concept of a biological switch, novel to iGEM community, by our team. Our team has combined the CRISPRi system with RNA regulators to achieve a toggle switch. The switch utilizes the catalytically inactive form of Cas9 (dCas9) to achieve targeted and reversible repression of genes via specific single-guide RNAs (sgRNAs). Alternatively, the transcription of antisense RNA (asRNAs) reverses the effect of the dCas9 modulated repression on the desired genes. This method of regulation would allow for the ability to fine-tune and easily customize the execution of highly complex genetic circuits.</p>
+<p>To bring our project to life, we have designed a circuit that drives the <i>Escherichia coli</i> (<i>E.coli</i>) strain DH5-alpha to alternately produce green fluorescent protein (GFP) and red fluorescent protein (RFP) depending on the inducer added to our system.</p>
+<h2> Proposed Toggle Switch Construct </h2>
+<img src="https://2019.igem.org/wiki/images/1/18/T--Hong_Kong_HKUST--Circuit.svg" width=800px>
 <!-- Add more about the biology of this part here