Detection and Manipulation of Live Antigen-expressing Cells Using Conditionally Stable Nanobodies

Overview

Journal Elife

Specialty Biology

Date 2016 May 21

PMID 27205882

Citations 47

Authors

Jonathan Cy Tang

Eugene Drokhlyansky

Behzad Etemad

Stephanie Rudolph

Binggege Guo

Sui Wang

Emily G Ellis

Jonathan Z Li

Constance L Cepko

Affiliations

Soon will be listed here.

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.

Citing Articles

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