Reporter

Part:BBa_K3380151

Designed by: Alexandru Popov   Group: iGEM20_Edinburgh   (2020-10-08)
Revision as of 09:26, 10 October 2020 by Alexandru popov (Talk | contribs) (Characterization)


DIR2s-Apt fluorescent RNA aptamer

DIR2s-Apt is a fluorescent RNA aptamer comprising a short ribonucleotide sequence that exhibits fluorescence upon binding to a specific fluorophore.

Characterization

The DIR2s-Apt fluorescent RNA aptamer was described by Tan et al. (2017).

The DIR2s-Apt folding prediction as simulated by [http://www.nupack.org// NUPACK] is shown below.

DIR2s-Apt Characteristics
Fluorophore DFHBI
Excitation wavelength 442 (nm)
Emission wavelength 503 (nm)
Quantum Yield 0.98
Extinction coefficient 26100 (M^-1 cm^-1)
Source [1]

DIR2s-Apt folding

Usage and Biology

It is usually used in live-cell imaging, however it can also be used in vitro. DIR2-Apt can be used for visualisation by gene fusion to the target of interest. Its fluorescent colour depends on the fluorophore it binds to. It can bind to DIR ( dimethylindole red) derivatives to give a red colour and OTB (oxazole thiazole blue) derivatives to give a blue colour. The emission spectra also depends on the bound fluorophore. The DIR-Profluorophore has an excitation wavelength of 600 (nm) and an emission wavelength of 658 (nm). The OTB-SO3fluorophore has an excitation wavelength of 380 (nm) and an emission wavelength of 421 (nm).

DIR2s-Apt promiscuity for multiple fluorophores (DIR and OTB) allows its colour change just by addition of different fluorophores. Its short length also allows it to be synthesized rather than being cloned using a plasmid.

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
//rna/aptamer
Parameters
None