Regulatory

Part:BBa_K4195071

Designed by: Xiaoping Yu   Group: iGEM22_XMU-China   (2022-09-26)
Revision as of 17:19, 10 October 2022 by 3489339288 (Talk | contribs)


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).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 348
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 250
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None