SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos

Claire E Dudley; Lotte van den Goor; Ann L Miller

doi:10.1016/j.xpro.2022.101622

SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos

STAR Protoc. 2022 Aug 18;3(3):101622. doi: 10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16.

Authors

Claire E Dudley¹, Lotte van den Goor², Ann L Miller³

Affiliations

¹ Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085, USA. Electronic address: dudleyc@umich.edu.
² Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085, USA.
³ Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085, USA; Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, MI 48109-1085, USA. Electronic address: annlm@umich.edu.

Abstract

Traditional fluorescent proteins exhibit limitations in brightness and photostability that hinder optimal characterization of the dynamic cellular behavior of proteins of interest. SNAP- and Halo-tagging are alternatives to traditional fluorescent protein tagging utilizing bright, stable chemical dyes, which may improve signal-to-noise ratio. However, there has been limited use of this approach in vivo in developing organisms. Here, we present a protocol for implementing SNAP- and Halo-tagging in gastrula-stage Xenopus laevis embryos for live confocal microscopy. For complete details on the use and execution of this protocol, please refer to Varadarajan et al. (2022).

Keywords: Cell biology; Microscopy; Model organisms.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Coloring Agents*
Microscopy, Confocal
Proteins*
Xenopus laevis

Substances

Coloring Agents
Proteins

Grants and funding

R01 GM112794/GM/NIGMS NIH HHS/United States