A genetically encoded sensor with improved fluorescence intensity for opioid detection at cellular resolution. Issue 81 (24th September 2021)

• Record Type: Journal Article
• Authors: Kroning, Kayla E.; Li, Mingcheng; Petrescu, D. Isabel; Wang, Wenjing
• Is Part Of: Chemical communications. Volume 57:Issue 81(2021)
• Page Start: 10560
• Page End: 10563
• Publication Date: 2021-09-24
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1039/d1cc04524e
• Languages: English
• ISSNs: 1359-7345

Abstract

Abstract : We engineered an improved opioid sensor, called M-SPOTIT2, which is 11x brighter than our previously engineered M-SPOTIT1.1. Scale bar, 20 μm. Abstract : The mu-opioid receptor (MOR) regulates the neuronal pathways involved in pain, reward, and respiration. To increase our understanding of MOR's roles in these pathways, there is a need to detect opioids at cellular resolution. Here, we engineered an improved opioid-sensor, called M-SPOTIT2, which is 11x brighter than our previously engineered M-SPOTIT1.1. We engineered M-SPOTIT2 by adding the amino acids YNSH, located near the fluorophore of the enhanced green fluorescent protein, to the circular permuted green fluorescent protein in M-SPOTIT2. M-SPOTIT2 is 11x brighter than our previously engineered M-SPOTIT1.1 in HEK293T cell culture and 2.7x brighter in neuronal culture. M-SPOTIT2 will potentially be useful for the detection of opioids in cell culture for drug screening and the detection of opioids at cellular resolution in animal tissues. By using M-SPOTIT2, researchers can gain more understanding about the mechanisms of addiction, respiratory suppression, and pain-modulation involved in opioid signaling.