Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
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DOI：
10.21769/bioprotoc.3413
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发表时间：
2019-11-05
期刊：
影响因子：
0.8
通讯作者：
Beatty, Kimberly E.
中科院分区：
文献类型：
--
作者：
Doh, Julia K.;Enns, Caroline A.;Beatty, Kimberly E.
遗传编码标签是用于细胞成像和多尺度细胞成像的有用工具。多功能相互作用肽（VIP）标签，如VIPER，是新的遗传编码的标签，可用于各种成像应用。VIP标签由卷曲螺旋异二聚体组成，其中一个肽用作遗传标签，另一个（“探针肽”）递送与成像相容的报告物。异源二聚体的形成是快速和特异的，允许蛋白质被选择性地标记用于活细胞和固定细胞成像。在这个生物协议中，我们包括一个详细的指南，用于实施VIPER技术，对活细胞上的受体和固定细胞中的细胞内靶点进行成像。该方案由概述VIPER的使用的两个其他生物方案补充（Doh等人，2019a和2019b）。
Genetically-encoded tags are useful tools for multicolor and multi-scale cellular imaging. Versatile Interacting Peptide (VIP) tags, such as VIPER, are new genetically-encoded tags that can be used in various imaging applications. VIP tags consist of a coiled-coil heterodimer, with one peptide serving as the genetic tag and the other ("probe peptide") delivering a reporter compatible with imaging. Heterodimer formation is rapid and specific, allowing proteins to be selectively labeled for live-cell and fixed-cell imaging. In this Bio-Protocol, we include a detailed guide for implementing the VIPER technology for imaging receptors on live cells and intracellular targets in fixed cells. This protocol is complemented by two other Bio-Protocols outlining the use of VIPER (Doh et al., 2019a and 2019b).