Use of genetically-encoded calcium indicators for live cell calcium imaging and localization in virus-infected cells

Methods : a Companion to Methods in Enzymology
Jacob L PerryJoseph M Hyser

Abstract

Calcium signaling is a ubiquitous and versatile process involved in nearly every cellular process, and exploitation of host calcium signals is a common strategy used by viruses to facilitate replication and cause disease. Small molecule fluorescent calcium dyes have been used by many to examine changes in host cell calcium signaling and calcium channel activation during virus infections, but disadvantages of these dyes, including poor loading and poor long-term retention, complicate analysis of calcium imaging in virus-infected cells due to changes in cell physiology and membrane integrity. The recent expansion of genetically-encoded calcium indicators (GECIs), including blue and red-shifted color variants and variants with calcium affinities appropriate for calcium storage organelles like the endoplasmic reticulum (ER), make the use of GECIs an attractive alternative for calcium imaging in the context of virus infections. Here we describe the development and testing of cell lines stably expressing both green cytoplasmic (GCaMP5G and GCaMP6s) and red ER-targeted (RCEPIAer) GECIs. Using three viruses (rotavirus, poliovirus and respiratory syncytial virus) previously shown to disrupt host calcium homeostasis, we show the GECI cel...Continue Reading

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Citations

Nov 22, 2015·Methods : a Companion to Methods in Enzymology·Karen Mossman
Aug 22, 2017·Frontiers in Physiology·Miranda A MathewsAndrew J Murray
Mar 20, 2020·Scientific Reports·Catherine Tomaro-DuchesneauRobert A Britton
Apr 1, 2020·American Journal of Physiology. Gastrointestinal and Liver Physiology·Melinda A EngevikJames Versalovic
Nov 21, 2020·Science·Alexandra L Chang-GrahamJoseph M Hyser
Jan 28, 2021·Micromachines·Csaba ForroGabriella Panuccio
Feb 1, 2021·The American Journal of Pathology·Faith D IhekweazuJames Versalovic

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