@article{cdd6a9d2ca9d42d8b0508b29693250d8,
title = "Detection and manipulation of live antigen-expressing cells using conditionally stable nanobodies",
abstract = "The ability to detect and/or manipulate specific cell populations based upon the presence of intracellular protein epitopes would enable many types of studies and applications. Protein binders such as nanobodies (Nbs) can target untagged proteins (antigens) in the intracellular environment. However, genetically expressed protein binders are stable regardless of antigen expression, complicating their use for applications that require cell-specificity. Here, we created a conditional system in which the stability of an Nb depends upon an antigen of interest. We identified Nb framework mutations that can be used to rapidly create destabilized Nbs. Fusion of destabilized Nbs to various proteins enabled applications in living cells, such as optogenetic control of neural activity in specific cell types in the mouse brain, and detection of HIV-infected human cells by flow cytometry. These approaches are generalizable to other protein binders, and enable the rapid generation of single-polypeptide sensors and effectors active in cells expressing specific intracellular epitopes.",
author = "Tang, {Jonathan C.Y.} and Eugene Drokhlyansky and Behzad Etemad and Stephanie Rudolph and Binggege Guo and Sui Wang and Ellis, {Emily G.} and Li, {Jonathan Z.} and Cepko, {Constance L.}",
note = "Funding Information: We thank S Zhao and C Wang of the Z He laboratory (Boston Children?s Hospital) for rAAV production (core service supported by grant NEI 5P30EY012196-17). We thank M Springer, B Huang, M Lichterfeld, E Scully, A Tsibris, D Kuritzkes and members of the Cepko/Tabin/Dymecki lab for input on the manuscript, and the Neurobiology Imaging Facility (supported by NINDS P30 Core Center grant NS072030) for consultation and instrument availability. We thank WG Regehr for research support. We thank the Dana Farber Flow Cytometry facility for assistance with FACS. We thank S Arber and Addgene for plasmids. In addition to direct funding, SR was funded by the US National Institutes of Health (R01 NS32405 and R01 NS092707 to WG Regehr). Howard Hughes Medical Institute Stephanie Rudolph, National Institutes of Health F32 NS087708 Binggege Guo, National Institutes of Health K08 AI100699 Jonathan Z Li, National Institutes of Health R21 AI114448 Constance L Cepko Publisher Copyright: {\textcopyright} Tang et al. Copyright: Copyright 2016 Elsevier B.V., All rights reserved.",
year = "2016",
month = may,
day = "20",
doi = "10.7554/eLife.15312",
language = "English (US)",
volume = "5",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
number = "MAY2016",
}