Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies

Olena S. Oliinyk, Mikhail Baloban, Charles L. Clark, Erin Carey, Sergei Pletnev, Axel Nimmerjahn, Vladislav V. Verkhusha

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Small near-infrared (NIR) fluorescent proteins (FPs) are much needed as protein tags for imaging applications. We developed a 17 kDa NIR FP, called miRFP670nano3, which brightly fluoresces in mammalian cells and enables deep-brain imaging. By exploring miRFP670nano3 as an internal tag, we engineered 32 kDa NIR fluorescent nanobodies, termed NIR-Fbs, whose stability and fluorescence strongly depend on the presence of specific intracellular antigens. NIR-Fbs allowed background-free visualization of endogenous proteins, detection of viral antigens, labeling of cells expressing target molecules and identification of double-positive cell populations with bispecific NIR-Fbs against two antigens. Applying NIR-Fbs as destabilizing fusion partners, we developed molecular tools for directed degradation of targeted proteins, controllable protein expression and modulation of enzymatic activities. Altogether, NIR-Fbs enable the detection and manipulation of a variety of cellular processes based on the intracellular protein profile.

Original languageEnglish (US)
Pages (from-to)740-750
Number of pages11
JournalNature Methods
Volume19
Issue number6
DOIs
StatePublished - Jun 2022

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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