TY - JOUR
T1 - Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies
AU - Oliinyk, Olena S.
AU - Baloban, Mikhail
AU - Clark, Charles L.
AU - Carey, Erin
AU - Pletnev, Sergei
AU - Nimmerjahn, Axel
AU - Verkhusha, Vladislav V.
N1 - Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2022/6
Y1 - 2022/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85130437154&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85130437154&partnerID=8YFLogxK
U2 - 10.1038/s41592-022-01467-6
DO - 10.1038/s41592-022-01467-6
M3 - Article
C2 - 35606446
AN - SCOPUS:85130437154
SN - 1548-7091
VL - 19
SP - 740
EP - 750
JO - Nature Methods
JF - Nature Methods
IS - 6
ER -