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mCherry contains a fluorescent protein isoform that interferes with its reporter function.

ABSTRACT: Fluorescent proteins are essential reporters in cell and molecular biology. Here, we found that red-fluorescent proteins possess an alternative translation initiation site that produces a short functional protein isoform in both prokaryotes and eukaryotes. The short isoform creates significant background fluorescence that biases the outcome of expression studies. In this study, we identified the short protein isoform, traced its origin, and determined the extent of the issue within the family of red fluorescent protein. Our analysis showed that the short isoform defect of the red fluorescent protein family may affect the interpretation of many published studies. We provided a re-engineered mCherry variant that lacks background expression as an improved tool for imaging and protein expression studies.

SUBMITTER: Fages-Lartaud M

PROVIDER: S-EPMC9395592 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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<i>mCherry</i> contains a fluorescent protein isoform that interferes with its reporter function.

Fages-Lartaud Maxime M Tietze Lisa L Elie Florence F Lale Rahmi R Hohmann-Marriott Martin Frank MF

Frontiers in bioengineering and biotechnology 20220809

Fluorescent proteins are essential reporters in cell and molecular biology. Here, we found that red-fluorescent proteins possess an alternative translation initiation site that produces a short functional protein isoform in both prokaryotes and eukaryotes. The short isoform creates significant background fluorescence that biases the outcome of expression studies. In this study, we identified the short protein isoform, traced its origin, and determined the extent of the issue within the family of ...[more]

PMID: 36017355

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mCherry contains a fluorescent protein isoform that interferes with its reporter function

Project description:Fluorescent proteins are essential reporters in cell biology and molecular biology. Here, we reveal that red-fluorescent proteins possess an alternative translation initiation site that produces a short functional protein isoform. The short isoform creates significant background fluorescence that biases the outcome of expression studies. Our investigation identifies the short protein isoform, traces its origin, and determines the extent of the issue within the family of red fluorescent protein. Our analysis shows that the short isoform defect of the red fluorescent protein family may affect the interpretation of many published studies. Finally, we provide a re-engineered mCherry variant that lacks background expression as an improved tool for imaging and protein expression studies.

2022-04-19 | PXD032954 | Pride

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