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Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.

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Expression of Fluorescent Proteins in Branchiostoma lanceolatum by mRNA Injection into Unfertilized Oocytes

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Introduction and Expression of Recombinant Genes in Ascidian Embryos.

Akira Hikosaka¹, Takehiro Kusakabe¹, Noriyuki Satoh¹

¹Department of Zoology, Faculty of Science, Kyoto University, Kyoto 606-01, Japan.

Development, Growth & Differentiation

|June 7, 2023

Summary

Exogenous gene expression in ascidian eggs was studied using two plasmids. Recombinant genes successfully expressed in various cell types, with one promoter directing muscle-specific expression.

Keywords:

(ascidian eggs gene expression microinjection recombinant genes

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Area of Science:

Developmental Biology
Molecular Biology
Genetics

Background:

Ascidian embryos are valuable models for studying gene expression during development.
Understanding exogenous gene expression is crucial for genetic manipulation and functional studies in ascidians.

Purpose of the Study:

To investigate the expression patterns of exogenous genes introduced into ascidian eggs.
To evaluate the efficacy of specific gene promoters in driving gene expression in different cell types.

Main Methods:

Microinjection of recombinant plasmids (pmiwZ with lac-Z, pHrMA4aCAT with CAT) into fertilized eggs of Ciona savignyi and Halocynthia roretzi.
Histochemical detection of β-galactosidase activity and anti-CAT antibody staining to assess protein expression.
Analysis of gene expression in various embryonic stages and cell types.

Main Results:

Microinjection of pmiwZ resulted in β-galactosidase expression in diverse cell types (epidermis, nervous system, muscle, etc.) across different embryonic stages.
pHrMA4aCAT introduced into Halocynthia eggs showed CAT expression specifically in muscle cells.
The 5' flanking region of the H. roretzi muscle actin gene (HrMA4a) contained sufficient regulatory sequences for appropriate spatial and temporal expression.

Conclusions:

Recombinant genes can be successfully expressed in ascidian embryos.
The HrMA4a promoter directs appropriate muscle-specific gene expression in ascidians.
Ascidian eggs serve as a viable system for studying gene expression and regulation.