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This novel strategy may provide a more physiologically relevant alternative to the traditional method of reporter cell line construction.
This method of reporter expression is commonly used in zebrafish research, and multiple lines within our lab using other transgenes (eg, SMN) made with this linked expression construct behave similarly.
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But generally they choose not to inquire too closely into the methods of reporters.
In some samples, it was confirmed that this method reproduces the results of relative promoter activity evaluated by the canonical method of CAT reporter assay using [C]-chloramphenicol.
This is an obvious improvement over the standard method of conjugating reporter dye precursors to the probes, which is limited to one fluorophore per target hybridization.
Our method allows generation of reporter cell lines which express simultaneously several proteins of interest, where each protein is labeled endogenously with a different fluorescent tag.
We used the efficient method of electroporation of reporter plasmids into developing retinal tissue to probe the activity of these TREs.
This method makes use of reporter constructs in which expression of EGFP is driven by DNA sequences from the locus of interest [ 23].
Therefore, we compared two methods of independent reporter coexpression and examined colocalization with induced connexin expression.
This allowed us to faithfully reproduce the natural expression pattern of ADGF-A in a manner offering maximal fidelity, especially when compared with more conventional methods of transgenic reporter fly generation that rely on random sites of genomic integration.
The paper contributes to the development of sound methods for the interpretation of reporter gene data, notably in the context of the reconstruction and validation of models of regulatory networks.
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