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ZeClinics genetic experts can investigate the result of overexpressing an endogenous gene or exogenous coding sequence (CDS) via mRNA injection in zebrafish embryos. It is a fast and very easy way to see if your gene of interest might have a phenotype (reverse genetic analysis), and warrant making a germline mutant/transgenic.
Transient overexpression experiments can be employed to mimic pathological conditions caused by aberrant protein expression and to quickly characterize the effect of this abnormal protein expression. Additionally, they can also be a useful strategy to restore the phenotypes observed in loss-of-function models.
Overexpression experiments will help establish a direct link between the biochemical function of a gene product and its role in vivo. Phenotyping is possible with good controls but the decreasing concentration of injected mRNA leads to sub-optimal KO usually by 4dpf.
Looking for a strategy to evaluate the effect of aberrant protein expression in pathological conditions?
Uniform protein overexpression at early developmental stages.High throughput screening of multiple candidate genesNon-invasive, in vivo readouts to detect gain-of-function-induced phenotypes at cell, tissue or organ levels.
Uniform protein overexpression at early developmental stages.
High throughput screening of multiple candidate genes
Non-invasive, in vivo readouts to detect gain-of-function-induced phenotypes at cell, tissue or organ levels.
To generate a model based on mRNA’s expression, zebrafish one-cell-stage embryos are injected with a solution containing the mRNA synthesized in vitro and encoding for the CDS of interest. During early stages of embryonic development, the plasmid can be imported in the nucleus and remain episomic. The exogenous CDS can be expressed by the promoter sequence included in the plasmid giving rise to F0 larvae transiently expressing the sequence of interest. The expression of the exogenous CDS can be associated with one fluorescent reporter to facilitate the phenotypic analysis of the models generated.
Results of the phenotypic analysis according to customers' interests and needs:
If you want more information about our mRNAinjection services or have any other questions,please contact our experts.