Difference between revisions of "Part:BBa K3453051"

Revision as of 22:28, 9 October 2020

Rosewood DmTrnL-UAA Toehold Switch 1.1

This part is a toehold switch sensor for sequence-based detection of rosewood. It targets a fragment of the TrnL-UAA gene of Dalbergia maritima var. pubescens (BBa_K3453060).

Usage and Biology

A toehold switch is an RNA–based device containing a ribosome binding site (RBS) and an ATG start codon embedded in a hairpin structure that blocks translation initiation [1]. The hairpin can be unfolded upon binding of a trigger RNA therefore exposing the RBS and permitting translation of the reporter protein.

This part is a toehold switch sensor that targets a fragment of the TrnL-UAA gene of Dalbergia maritima var. pubescens (BBa_K3453060). It was designed using the web tool developed by To et al. [2] and follows the architecture of the Series B of toehold switch sensors for Zika virus detection [3] and of the BioBits™ toeholds [4].

Its corresponding trigger sequence is BBa_K3453061.

The functionality of this part was tested using sfGFP-LVAtag (BBa_K2675006) as a reporter. The expression was controlled by the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000) in the composite part BBa_K3453151.

This part proved not to be functional: a readable output was generated even in the absence of rosewood RNA.

Full results are available on the BBa_K3453151 page in the registry.

References

[1] Green AA, Silver PA, Collins JJ, Yin P. Toehold switches: de-novo-designed regulators of gene expression. Cell (2014) 159, 925-939.

[2] To AC, Chu DH, Wang AR, Li FC, Chiu AW, Gao DY, Choi CHJ, Kong SK, Chan TF, Chan KM, Yip KY. A comprehensive web tool for toehold switch design. Bioinformatics (2018) 34, 2862-2864.

[3] Pardee K, Green AA, Takahashi MK, Braff D, Lambert G, Lee JW, Ferrante T, Ma D, Donghia N, Fan M, Daringer NM, Bosch I, Dudley DM, O'Connor DH, Gehrke L, Collins JJ. Rapid, low-cost detection of Zika virus using programmable biomolecular components. Cell (2016) 165, 1255-1266.

[4] Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ. BioBits™ Explorer: A modular synthetic biology education kit. Sci Adv (2018) 4, eaat5105.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 7: / Line 7: @@
 ===Usage and Biology===
-A toehold switch is an RNA–based device containing a ribosome binding site (RBS) embedded in a hairpin structure that blocks translation initiation [1]. The hairpin can be unfolded upon binding of a trigger RNA therefore exposing the RBS and permitting translation of the reporter protein.
+A toehold switch is an RNA–based device containing a ribosome binding site (RBS) and an ATG start codon embedded in a hairpin structure that blocks translation initiation [1]. The hairpin can be unfolded upon binding of a trigger RNA therefore exposing the RBS and permitting translation of the reporter protein.
 This part is a toehold switch sensor that targets a fragment of the TrnL-UAA gene of ''Dalbergia maritima'' var. ''pubescens'' ([[Part:BBa_K3453060|BBa_K3453060]]).