Cell-Type-Specific Gene Inactivation and In Situ Restoration via Recombinase-Based Flipping of Targeted Genomic Region
| Autor: | Xue Liu, Yidan Xu, Peiyuan Mu, Yun Shi, Liang Ma, Miao He, Yubin Zhang, Ling Gong, Tianrui Zhang, Jinyun Wu, Jingwen Gan, Hongzhi Liu |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Male Methyl-CpG-Binding Protein 2 Movement Rett syndrome Computational biology Biology MECP2 03 medical and health sciences Mice 0302 clinical medicine Recombinase medicine Rett Syndrome Animals Allele Gene X chromosome Research Articles Germ-Line Mutation Regulation of gene expression Neurons Integrases General Neuroscience medicine.disease Phenotype Mice Inbred C57BL 030104 developmental biology DNA Nucleotidyltransferases Gene Targeting 030217 neurology & neurosurgery |
| Zdroj: | J Neurosci |
| Popis: | Conditional gene inactivation and restoration are powerful tools for studying gene functions in the nervous system and for modeling neuropsychiatric diseases. The combination of the two is necessary to interrogate specific cell types within defined developmental stages. However, very few methods and animal models have been developed for such purpose. Here we present a versatile method for conditional gene inactivation and in situ restoration through reversibly inverting a critical part of its endogenous genomic sequence by Cre- and Flp-mediated recombinations. Using this method, we generated a mouse model to manipulate Mecp2, an X-linked dosage-sensitive gene whose mutations cause Rett syndrome. Combined with multiple Cre- and Flp-expressing drivers and viral tools, we achieved efficient and reliable Mecp2 inactivation and restoration in the germline and several neuronal cell types, and demonstrated phenotypic reversal and prevention on cellular and behavioral levels in male mice. This study not only provides valuable tools and critical insights for Mecp2 and Rett syndrome, but also offers a generally applicable strategy to decipher other neurologic disorders. SIGNIFICANCE STATEMENT Studying neurodevelopment and modeling neurologic disorders rely on genetic tools, such as conditional gene regulation. We developed a new method to combine conditional gene inactivation and restoration on a single allele without disturbing endogenous expression pattern or dosage. We applied it to manipulate Mecp2, a gene residing on X chromosome whose malfunction leads to neurologic disease, including Rett syndrome. Our results demonstrated the efficiency, specificity, and versatility of this new method, provided valuable tools and critical insights for Mecp2 function and Rett syndrome research, and offered a generally applicable strategy to investigate other genes and genetic disorders. |
| Databáze: | OpenAIRE |
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