RNA

Part:BBa_K3453042

Designed by: Maeva Cherriere   Group: iGEM20_Evry_Paris-Saclay   (2020-10-04)


Rosewood DmRbcL Trigger 1.2

This part is a trigger for sequence-based detection of rosewood. It is a fragment of the RbcL gene of Dalbergia maritima var. pubescens (BOLD Acc. n° MADA155-14, MADA063-14, MADA062-14, MADA158-14, MADA065-14, MADA104-14, MADA023-14, MADA071-14, MADA110-14) [1].

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 [2]. The hairpin can be unfolded upon binding of a trigger RNA thereby exposing the RBS and the ATG start codon permitting translation of the reporter protein (Figure 1).

T--Evry Paris-Saclay--toehold-switch.png

Figure 1. Toehold switches principle.

This part is a trigger for the Rosewood DmRbcL Toehold Switch 1.2 (BBa_K3453032) and was designed using the web tool developed by To et al. [3]. It also contains the switch binding site for the Rosewood DmRbcL Toehold Switch 1.1 (BBa_K3453031).

The functionality of this part was tested under controlled of the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000) in the composite part BBa_K3453142.

This part proved to be functional: the transcribed rosewood RNA was able to bind to the Rosewood DmRbcL Toehold Switch 1.1 and a readable output was generated. However, the Rosewood DmRbcL Toehold Switch 1.2 turned out to be not functional and the interaction with this switch could not be assessed.

Full results are available on the BBa_K3453131 and BBa_K3453132 pages in the registry.

References

[1] Hassold S, Lowry PP 2nd, Bauert MR, Razafintsalama A, Ramamonjisoa L, Widmer A. DNA barcoding of Malagasy rosewoods: towards a molecular identification of CITES-listed Dalbergia species. PLoS One (2016) 11, e0157881.

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

[3] 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.

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]


[edit]
Categories
Parameters
None