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Using <i>i</i>-GONAD for Cell-Type-Specific and Systematic Analysis of Developmental Transcription Factors In Vivo
oleh: Christoph Wiegreffe, Simon Ehricke, Luisa Schmid, Jacqueline Andratschke, Stefan Britsch
Format: | Article |
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Diterbitkan: | MDPI AG 2023-09-01 |
Deskripsi
Transcription factors (TFs) regulate gene expression via direct DNA binding together with cofactors and in chromatin remodeling complexes. Their function is thus regulated in a spatiotemporal and cell-type-specific manner. To analyze the functions of TFs in a cell-type-specific context, genome-wide DNA binding, as well as the identification of interacting proteins, is required. We used <i>i</i>-GONAD (improved genome editing via oviductal nucleic acids delivery) in mice to genetically modify TFs by adding fluorescent reporter and affinity tags that can be exploited for the imaging and enrichment of target cells as well as chromatin immunoprecipitation and pull-down assays. As proof-of-principle, we showed the functional genetic modification of the closely related developmental TFs, <i>Bcl11a</i> and <i>Bcl11b</i>, in defined cell types of newborn mice. <i>i</i>-GONAD is a highly efficient procedure for modifying TF-encoding genes via the integration of small insertions, such as reporter and affinity tags. The novel <i>Bcl11a</i> and <i>Bcl11b</i> mouse lines, described in this study, will be used to improve our understanding of the Bcl11 family’s function in neurodevelopment and associated disease.