Source 1primary paper2018PLoS ONE
Toolkit/humanized mNeonGreen-3xFLAG
humanized mNeonGreen-3xFLAG
Also known as: 3xFLAG-tagged mNeonGreen, mNeonGreen-3xFLAG
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Humanized mNeonGreen-3xFLAG is a mammalian expression construct encoding a human codon-optimized mNeonGreen fluorescent protein fused at the carboxyl terminus to a 3xFLAG epitope tag. It enables fluorescent visualization together with anti-FLAG antibody-based detection in mammalian cells.
Usefulness & Problems
Why this is useful
This construct is useful for experiments that require both live-cell fluorescent reporting and immunological detection of the same expressed protein product. The humanized mNeonGreen backbone was reported to increase fluorescent intensity in HEK293 cells relative to the original mNeonGreen, and the 3xFLAG fusion was reported to be well recognized by anti-FLAG-M2 antibody.
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 single mammalian expression reagent that combines bright fluorescent labeling with a standard epitope tag for antibody-based detection. The reported human codon optimization specifically targets reduced performance of the original mNeonGreen in mammalian cells.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Techniques
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 reported construct is an expression vector encoding humanized mNeonGreen with 3xFLAG tags at its carboxyl terminus for mammalian expression. Evidence supports use in HEK293 cells, and a separate humanized mNeonGreen variant bearing a mitochondria-targeting signal showed mitochondrial distribution, indicating compatibility of the humanized fluorophore with additional targeting sequences.
The supplied evidence is limited to a single source and does not report broad validation across multiple mammalian cell types, organisms, or assay formats. The evidence also does not quantify effects of the 3xFLAG fusion on brightness, maturation, photostability, or protein localization beyond a separate mitochondria-targeted humanized mNeonGreen example.
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-3xflag
We further generated an expression vector of humanized mNeonGreen with 3xFLAG tags at its carboxyl terminus
Source:
immunological detectabilitysupports
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.
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
In the cited study, humanized mNeonGreen showed higher fluorescent intensity than the original mNeonGreen in HEK293 cells. The carboxyl-terminal 3xFLAG-tagged version was recognized well by anti-FLAG-M2 antibody, supporting compatibility with FLAG-based immunodetection workflows.
Compared with joining proteins in creative ways
humanized mNeonGreen-3xFLAG and joining proteins in creative ways address a similar problem space.
Shared frame: same top-level item type
Compared with Nano-lantern
humanized mNeonGreen-3xFLAG and Nano-lantern address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Compared with tethered PEs
humanized mNeonGreen-3xFLAG and tethered PEs address a similar problem space.
Shared frame: same top-level item type
Ranked Citations
- 1.