Interdisciplinary Bioengineering Graduate Program, Georgia Institute of Technology, Atlanta, GA 30332, USA.
School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.
G3 (Bethesda). 2023 Sep 30;13(10). doi: 10.1093/g3journal/jkad183.
In living organisms, changes in calcium flux are integral to many different cellular functions and are especially critical for the activity of neurons and myocytes. Genetically encoded calcium indicators (GECIs) have been popular tools for reporting changes in calcium levels in vivo. In particular, GCaMPs, derived from GFP, are the most widely used GECIs and have become an invaluable toolkit for neurophysiological studies. Recently, new variants of GCaMP, which offer a greater variety of temporal dynamics and improved brightness, have been developed. However, these variants are not readily available to the Caenorhabditis elegans research community. This work reports a set of GCaMP6 and jGCaMP7 reporters optimized for C. elegans studies. Our toolkit provides reporters with improved dynamic range, varied kinetics, and targeted subcellular localizations. Besides optimized routine uses, this set of reporters is also well suited for studies requiring fast imaging speeds and low magnification or low-cost platforms.
在生物体中，钙流的变化是许多不同细胞功能的组成部分，特别是对神经元和肌细胞的活性至关重要。遗传编码的钙指示剂（GECIs）一直是报告体内钙水平变化的流行工具。特别是，源自 GFP 的 GCaMPs 是应用最广泛的 GECIs，已成为神经生理学研究的宝贵工具包。最近，开发了具有更大时间动态和更高亮度的新型 GCaMP 变体。然而，这些变体并不容易被秀丽隐杆线虫研究界所获得。本工作报道了一组针对秀丽隐杆线虫研究优化的 GCaMP6 和 jGCaMP7 报告器。我们的工具包提供了具有改进的动态范围、不同动力学和靶向亚细胞定位的报告器。除了优化的常规用途外，这套报告器也非常适合需要快速成像速度、低放大倍数或低成本平台的研究。
