Difference between revisions of "Part:BBa K4195071"

 
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<partinfo>BBa_K4195071 short</partinfo>
 
<partinfo>BBa_K4195071 short</partinfo>
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===Biology===
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This sequence is the first part of the guide.
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<br>
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<b>Ribozyme ENabled Detection of RNA (RENDR)</b>
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===Usage and Biology===
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<br>
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RENDR is a high-performing, plug-and-play RNA-sensing platform[1]. RENDR utilizes a split variant of the ''Tetrahymena thermophila'' ribozyme by synthetically splitting it into two non-functional fragments (Fig. 1). Two fragments are each appended with designed RNA guide sequences, which can interact with the RNA input of interest. The split ribozyme is then inserted within a desired gene output. When bound with the RNA input, two transcribed split ribozyme fragments are triggered to self-splice and thus the intact transcript of the protein output will form.<br/>
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[[File:T--XMU-China--comp.png|400px]]<br/>
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<span class='h3bb'>Sequence and Features</span>
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'''Fig. 1 Schematic illustration of RENDR.'''<br/>
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<partinfo>BBa_K4195071 SequenceAndFeatures</partinfo>
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===Usage and Design===
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We replicate the circuit used in the literature as reference, and separately designed two split ribozymes as different parts <partinfo>BBa_K4195074</partinfo> and <partinfo>BBa_K4195071</partinfo>. The combined one (<partinfo>BBa_K4195182</partinfo>) was assembled into the vector pSB1C3 by standard BioBrick assembly. After transcription, two RNA guides can interact with each other. <br/>
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===Characterization===
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Agarose Gel Electrophoresis<br/>
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===Functional Parameters===
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<partinfo>BBa_K4195182</partinfo> was assembled into the vector pSB1C3 by standard BioBrick assembly. The constructed plasmids were transformed into ''E. coli ''BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.<br/>
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<partinfo>BBa_K4195071 parameters</partinfo>
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[[File:T--XMU-China--comp g1β.png|400px]]<br/>
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Fig. 2 The result of colony PCR. Plasmid pSB1C3<br/>
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===Reference===
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[1]L. Gambill ''et al.'', https://www.biorxiv.org/content/10.1101/2022.01.12.476080v1 (2022).

Revision as of 16:14, 10 October 2022


comp_g1βF

Biology

This sequence is the first part of the guide.
Ribozyme ENabled Detection of RNA (RENDR)
RENDR is a high-performing, plug-and-play RNA-sensing platform[1]. RENDR utilizes a split variant of the Tetrahymena thermophila ribozyme by synthetically splitting it into two non-functional fragments (Fig. 1). Two fragments are each appended with designed RNA guide sequences, which can interact with the RNA input of interest. The split ribozyme is then inserted within a desired gene output. When bound with the RNA input, two transcribed split ribozyme fragments are triggered to self-splice and thus the intact transcript of the protein output will form.
T--XMU-China--comp.png
Fig. 1 Schematic illustration of RENDR.

Usage and Design

We replicate the circuit used in the literature as reference, and separately designed two split ribozymes as different parts BBa_K4195074 and BBa_K4195071. The combined one (BBa_K4195182) was assembled into the vector pSB1C3 by standard BioBrick assembly. After transcription, two RNA guides can interact with each other.

Characterization

Agarose Gel Electrophoresis
BBa_K4195182 was assembled into the vector pSB1C3 by standard BioBrick assembly. The constructed plasmids were transformed into E. coli BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.
T--XMU-China--comp g1β.png
Fig. 2 The result of colony PCR. Plasmid pSB1C3

Reference

[1]L. Gambill et al., https://www.biorxiv.org/content/10.1101/2022.01.12.476080v1 (2022).