Tag

Part:BBa_K4375008

Designed by: Zska Csorba   Group: iGEM22_ELTE   (2022-09-28)


miRFP670 nano

miRFP670nano is a near-infrared fluorescent protein published in 2019, derived from Nostoc punctiforme. It is special because it harbors an unusually small size. (17 kDa)


Usage and Biology

miRFP670nano is a small, 17 kDa, monomeric near-infrared fluorescent protein (NIR FP). Its excitation is at 645 nm and its emission is at 670 nm. NIR fluorescent proteins are useful in biological applications like deep tissue imaging as it has a better penetrative trait than visible light which is scattered by lipids. Most of them are derived from bacterial phytochrome photoreceptors as monomers because these have absorbance in NIR range spectra. miRFP670nano is derived from cyanobacteriochromes, which usually bind phycocyanobilins found in plants, however, this protein was engineered by various mutations to bind biliverdin. The protein has a GAF domain (cGMP phosphodiesterase-adenylate cyclase-FhlA), which has a major role in light perception in phytochromes. It has a monomeric state, is small, has high stability, and can be easily used as a fused tag. It also has a brightness that exceeds bacterial phytochrome photoreceptor-derived NIR fluorescent proteins even in mammalian cells.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 52
    Illegal PstI site found at 310
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 52
    Illegal PstI site found at 310
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 52
    Illegal PstI site found at 310
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 52
    Illegal PstI site found at 310
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Oliinyk, O. S.; Shemetov, A. A.; Pletnev, S.; Shcherbakova, D. M.; Verkhusha, V. V. Smallest Near-Infrared Fluorescent Protein Evolved from Cyanobacteriochrome as Versatile Tag for Spectral Multiplexing. Nature Communications 2019, 10 (1). https://doi.org/10.1038/s41467-018-08050-8.

Oliinyk, O.; Chernov, K.; Verkhusha, V. Bacterial Phytochromes, Cyanobacteriochromes and Allophycocyanins as a Source of Near-Infrared Fluorescent Probes. International Journal of Molecular Sciences 2017, 18 (8), 1691. https://doi.org/10.3390/ijms18081691.


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