How to Increase Brightness of Near-Infrared Fluorescent Proteins in Mammalian Cells

Anton A. Shemetov, Olena S. Oliinyk, Vladislav V. Verkhusha

Research output: Contribution to journalArticlepeer-review

48 Scopus citations


Numerous near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial photoreceptors but lack of their systematic comparison makes researcher's choice rather difficult. Here we evaluated side-by-side several modern NIR FPs, such as blue-shifted smURFP and miRFP670, and red-shifted mIFP and miRFP703. We found that among all NIR FPs, miRFP670 had the highest fluorescence intensity in various mammalian cells. For instance, in common HeLa cells miRFP703, mIFP, and smURFP were 2-, 9-, and 53-fold dimmer than miRFP670. Either co-expression of heme oxygenase or incubation of cells with heme precursor weakly affected NIR fluorescence, however, in the latter case elevated cellular autofluorescence. Exogenously added chromophore substantially increased smURFP brightness but only slightly enhanced brightness of other NIR FPs. mIFP showed intermediate, while monomeric miRFP670 and miRFP703 exhibited high binding efficiency of endogenous biliverdin chromophore. This feature makes them easy to use as GFP-like proteins for spectral multiplexing with FPs of visible range.

Original languageEnglish (US)
Pages (from-to)758-766.e3
JournalCell Chemical Biology
Issue number6
StatePublished - Jun 22 2017


  • 5-ALA
  • IFP
  • biliverdin
  • firefly luciferase
  • heme oxygenase
  • iRFP
  • in vivo imaging
  • near infrared
  • phytochrome
  • smURFP

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry


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