Rapid promoter analysis in developing mouse brain and genetic labeling of young neurons by doublecortin-DsRed-express.
Characterization of neural promoter/enhancers is essential for understanding gene regulation during brain development and provides useful genetic tools. However, it relies on the use of transgenic mice. We report a method for the rapid in vivo analysis of neural promoter/enhancers in the developing mouse brain and its application in the isolation of the doublecortin (DCX) promoter/enhancer for genetic labeling of young neurons. In the present study, we demonstrated that reporter genes introduced into the developing mouse cerebral cortex by in utero electroporation can achieve promoter/enhancer-specific patterns of expression. We used the in utero electroporation system to isolate a genomic fragment of the doublecortin gene that can direct reporter expression faithful to doublecortin in young neurons of the cerebral cortex. Finally, we showed that the DCX promoter identified via electroporation could reproduce doublecortin expression in the entire central nervous system in DCX-DsRed-express transgenic mice. The results of our study provide a convenient, reliable, and rapid method for in vivo analysis of neural promoter/enhancers in the developing mouse brain.
Wang
Xiuyun
X
Division of Neurosciences, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, CA 91010, USA.
Qiu
Runxiang
R
Tsark
Walter
W
Lu
Qiang
Q
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D001921:Brain / Q000196:embryology*; D019070:Cell Lineage / Q000235:genetics*; D002540:Cerebral Cortex / Q000196:embryology; D051360:Cyclin-Dependent Kinase 5 / Q000235:genetics; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D018274:Electroporation; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D016678:Genome / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013194:Staining and Labeling / Q000379:methods; D014162:Transfection
C000714847:Dcx protein, mouse; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides; C413663:fluorescent protein 583; D051360:Cyclin-Dependent Kinase 5; C495906:Cdk5 protein, mouse