A genetically encoded sensor with improved fluorescence intensity for opioid detection at cellular resolution

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, whic...

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Bibliographic Details
Published in:Chemical communications (Cambridge, England) England), 2021-10, Vol.57 (81), p.156-1563
Main Authors: Kroning, Kayla E, Li, Mingcheng, Petrescu, D. Isabel, Wang, Wenjing
Format: Article
Language:English
Subjects:
Amino acids
Cell culture
Cells, Cultured
Fluorescence
Genetic code
Green Fluorescent Proteins - genetics
HEK293 Cells
Humans
Models, Molecular
Narcotics
Pain
Proteins
Receptors, Opioid, mu - analysis
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Summary: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. We engineered an improved opioid sensor, called M-SPOTIT2, which is 11x brighter than our previously engineered M-SPOTIT1.1. Scale bar, 20 μm.
ISSN:1359-7345
1364-548X
DOI:10.1039/d1cc04524e