PUBLICATION
Genetically encoded affinity reagents are a toolkit for visualizing and manipulating endogenous protein function in vivo
- Authors
- Boswell, C.W., Hoppe, C., Sherrard, A., Miao, L., Kojima, M.L., Martino, P., Zhao, N., Stasevich, T.J., Nicoli, S., Giraldez, A.J.
- ID
- ZDB-PUB-250702-20
- Date
- 2025
- Source
- Nature communications 16: 55035503 (Journal)
- Registered Authors
- Boswell, Curtis, Giraldez, Antonio, Nicoli, Stefania
- Keywords
- none
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems*/genetics
- Epitopes/genetics
- Epitopes/metabolism
- Gene Knock-In Techniques*/methods
- Single-Chain Antibodies/genetics
- Single-Chain Antibodies/metabolism
- Single-Domain Antibodies/genetics
- Single-Domain Antibodies/metabolism
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 40595661 Full text @ Nat. Commun.
Citation
Boswell, C.W., Hoppe, C., Sherrard, A., Miao, L., Kojima, M.L., Martino, P., Zhao, N., Stasevich, T.J., Nicoli, S., Giraldez, A.J. (2025) Genetically encoded affinity reagents are a toolkit for visualizing and manipulating endogenous protein function in vivo. Nature communications. 16:55035503.
Abstract
Probing endogenous protein localization and function in vivo remains challenging due to laborious gene targeting and monofunctional alleles. Here, we develop a multifunctional and adaptable toolkit based on genetically encoded affinity reagents (GEARs). GEARs use small epitopes recognized by nanobodies and single chain variable fragments to enable fluorescent visualization, manipulation and degradation of protein targets in vivo. Furthermore, we outline a CRISPR/Cas9-based epitope tagging pipeline to demonstrate its utility for producing knock-in alleles that have broad applications. We use GEARs to examine the native behavior of the pioneer transcription factor Nanog and the planar cell polarity protein Vangl2 during early zebrafish development. Together, this toolkit provides a versatile system for probing and perturbing endogenous protein function while circumventing challenges associated with conventional gene targeting and is broadly available to the model organism community.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping