Ding Jimmy, Peng Lucinda, Moon Sihoon, Lee Hyun Jee, Patel Dhaval S, Lu Hang
These authors contributed equally.
Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA.
bioRxiv. 2023 Mar 8:2023.03.06.531342. doi: 10.1101/2023.03.06.531342.
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 particular, GCaMP, 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 research community. This work reports a set of GCaMP6 and jGCaMP7 reporters optimized for studies. Our toolkit provides reporters with improved dynamic range, varied kinetics, and targeted subcellular localizations. Besides optimized routine uses, this set of reporters are also well-suited for studies requiring fast imaging speeds and low magnification or low-cost platforms.
在活生物体中,钙通量的变化是许多不同细胞功能所不可或缺的,对神经元和心肌细胞的活动尤为关键。基因编码钙指示剂(GECIs)一直是用于报告钙水平变化的常用工具。特别是源自绿色荧光蛋白(GFP)的绿色荧光钙指示剂(GCaMP),是使用最为广泛的GECIs,已成为神经生理学研究中不可或缺的工具集。最近,已开发出具有更多种时间动态特性且亮度有所提高的新型GCaMP变体。然而,这些变体尚未在研究界广泛使用。这项工作报告了一组针对研究进行优化的GCaMP6和jGCaMP7报告基因。我们的工具集为报告基因提供了更高的动态范围、多样的动力学特性以及靶向亚细胞定位。除了优化的常规用途外,这组报告基因还非常适合需要快速成像速度以及低放大倍数或低成本平台的研究。