Source 1primary paper2018PLoS ONE
Toolkit/humanized mNeonGreen
humanized mNeonGreen
Also known as: human-optimized cDNA encoding mNeonGreen
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Humanized mNeonGreen is a human codon-optimized cDNA construct encoding the fluorescent protein mNeonGreen for mammalian expression. In HEK293 cells, it produced higher fluorescent intensity than the original mNeonGreen, and derivative constructs supported anti-FLAG detection and mitochondrial targeting.
Usefulness & Problems
Why this is useful
This construct is useful as a mammalian fluorescent reporter with improved signal relative to the original mNeonGreen in HEK293 cells. Reported derivative plasmids, including 3xFLAG-tagged and mitochondria-targeted versions, extend its use to immunodetection and subcellular localization studies in mammalian cells.
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 problem of suboptimal performance of the original mNeonGreen coding sequence in human cells by providing a human-optimized expression construct. The reported derivatives also help couple fluorescence to antibody-based detection and organelle-specific localization.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
epitope tag-based immunodetectionfluorescencesubcellular targeting via 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: sensor
The tool was generated by synthesizing a human-optimized cDNA encoding mNeonGreen and cloning it into an expression plasmid. Reported construct engineering included fusion to a 3xFLAG epitope tag and addition of a mitochondria-targeting signal for mammalian cell applications.
The evidence is limited to a single 2018 PLoS ONE study and explicitly reported validation in HEK293 cells. No broader cross-cell-type, in vivo, photophysical, or quantitative benchmarking details are provided in the supplied evidence.
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 source2 linked approval claimsfirst-pass slug humanized-mneongreen
we synthesized a human-optimized cDNA encoding mNeonGreen and generated an expression plasmid for humanized mNeonGreen
Source:
expression optimizationsupports
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.
Source:
utility statementsupports
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.
Source:
Comparisons
Source-backed strengths
The key reported strength is higher fluorescent intensity than the original mNeonGreen in HEK293 cells. A 3xFLAG-tagged version was recognized well by anti-FLAG-M2 antibody, and a mitochondria-targeted version showed mitochondrial distribution, supporting construct versatility in mammalian cell studies.
Compared with FMN fluorescence
humanized mNeonGreen and FMN fluorescence address a similar problem space.
Shared frame: shared mechanisms: fluorescence
Strengths here: looks easier to implement in practice.
Compared with humanized mNeonGreen-3xFLAG
humanized mNeonGreen and humanized mNeonGreen-3xFLAG address a similar problem space.
Shared frame: same top-level item type
Compared with mitochondria-targeted humanized mNeonGreen
humanized mNeonGreen and mitochondria-targeted humanized mNeonGreen address a similar problem space.
Shared frame: same top-level item type
Ranked Citations
- 1.