Source 1primary paper2018PLoS ONE
Toolkit/mitochondria-targeted humanized mNeonGreen
mitochondria-targeted humanized mNeonGreen
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Mitochondria-targeted humanized mNeonGreen is a mammalian expression construct in which a humanized mNeonGreen fluorescent protein is fused to a mitochondria-targeting signal. In the cited PLoS ONE study, this fusion showed mitochondrial distribution in mammalian cells, indicating utility for fluorescent visualization of mitochondria.
Usefulness & Problems
Why this is useful
This construct is useful for labeling mitochondria in mammalian cells with a bright fluorescent protein variant optimized for Homo sapiens expression. The underlying study also reported that humanized mNeonGreen had higher fluorescent intensity than the original mNeonGreen in HEK293 cells, supporting its use when stronger signal is desired.
Source:
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Problem solved
It addresses the need for a mitochondria-localized fluorescent reporter that is compatible with mammalian expression and subcellular imaging. The humanization step also addresses reduced fluorescence performance of the original mNeonGreen in mammalian cells, as assessed in HEK293 cells.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
fluorescent protein emission for subcellular visualizationorganelle targeting via a mitochondria-targeting signalTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: actuator
Implementation consists of fusing a mitochondria-targeting signal to a humanized mNeonGreen coding sequence for mammalian expression. The source study additionally supports that humanized mNeonGreen performs well in HEK293 cells and that a 3xFLAG-tagged version can be detected by anti-FLAG-M2 antibody, but the supplied evidence does not provide construct architecture details beyond the targeting fusion.
Evidence for this specific construct is limited to a report of mitochondrial distribution, without quantitative localization metrics, photophysical characterization, or functional perturbation analysis. Validation appears to come from a single study, and the supplied evidence does not specify the targeting peptide sequence, cell types used for the mitochondrial localization experiment, or performance relative to other mitochondrial markers.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
Humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells.
The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen.
fluorescent intensity fold change 1.4 fold
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
3xFLAG-tagged humanized mNeonGreen was recognized well by anti-FLAG-M2 antibody.
The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Plasmids expressing humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells.
These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.
Approval Evidence
1 source1 linked approval claimfirst-pass slug mitochondria-targeted-humanized-mneongreen
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen
Source:
subcellular localizationsupports
Humanized mNeonGreen bearing a mitochondria-targeting signal showed mitochondrial distribution.
The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen.
Source:
Comparisons
Source-backed strengths
The available evidence shows correct mitochondrial distribution when a mitochondria-targeting signal is appended to humanized mNeonGreen. Separate data from the same study indicate that humanized mNeonGreen is brighter than the original mNeonGreen in HEK293 cells and that a 3xFLAG-tagged version is readily detected by anti-FLAG-M2 antibody.
Compared with humanized mNeonGreen
mitochondria-targeted humanized mNeonGreen and humanized mNeonGreen address a similar problem space.
Shared frame: same top-level item type
Ranked Citations
- 1.