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In situ hybridization probe sequences for mouse Alx3, Fgf8, Msx1, Shh, Six3

Citation

Dinsmore, Colin (2021), In situ hybridization probe sequences for mouse Alx3, Fgf8, Msx1, Shh, Six3, Dryad, Dataset, https://doi.org/10.5061/dryad.mgqnk9916

Abstract

Serum response factor (SRF) is an essential transcription factor that influences many cellular processes including cell proliferation, migration, and differentiation. SRF directly regulates and is required for immediate early gene (IEG) and actin cytoskeleton-related gene expression. SRF coordinates these competing transcription programs through discrete sets of cofactors, the Ternary Complex Factors (TCFs) and Myocardin Related Transcription Factors (MRTFs). The relative contribution of these two programs to in vivo SRF activity and mutant phenotypes is not fully understood. To study how SRF utilizes its cofactors during development, we generated a knock-in SrfaI allele in mice harboring point mutations that disrupt SRF-MRTF-DNA complex formation but leave SRF-TCF activity unaffected. Homozygous SrfaI/aI mutants die at E10.5 with notable cardiovascular phenotypes, and neural crest conditional mutants succumb at birth to defects of the cardiac outflow tract but display none of the craniofacial phenotypes associated with complete loss of SRF in that lineage. Our studies further support an important role for MRTF mediating SRF function in cardiac neural crest and suggest new mechanisms by which SRF regulates transcription during development.

Methods

DNA plasmids encoding in situ hybridization probes were shared with the Soriano lab. These were subject to Sanger sequencing using T7 and T3 primers. The resulting sequences were assembled into annotated .dna files. When a plasmid backbone was known or could be identified through the sequencing reads, the entire plasmid was constructed.

Usage Notes

These can be read using free SnapGene software.