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Dev Dyn 2017 Sep 01;2469:657-669. doi: 10.1002/dvdy.24532.
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Xenopus pitx3 target genes lhx1 and xnr5 are identified using a novel three-fluor flow cytometry-based analysis of promoter activation and repression.

Hooker LN , Smoczer C , Abbott S , Fakhereddin M , Hudson JW , Crawford MJ .

BACKGROUND: Pitx3 plays a well understood role in directing development of lens, muscle fiber, and dopaminergic neurons; however, in Xenopus laevis, it may also play a role in early gastrulation and somitogenesis. Potential downstream targets of pitx3 possess multiple binding motifs that would not be readily accessible by conventional promoter analysis. RESULTS: We isolated and characterized pitx3 target genes lhx1 and xnr5 using a novel three-fluor flow cytometry tool that was designed to dissect promoters with multiple binding sites for the same transcription factor. This approach was calibrated using a known pitx3 target gene, Tyrosine hydroxylase. CONCLUSIONS: We demonstrate how flow cytometry can be used to detect gene regulatory changes with exquisite precision on a cell-by-cell basis, and establish that in HEK293 cells, pitx3 directly activates lhx1 and represses xnr5. Developmental Dynamics 246:657-669, 2017. © 2017 Wiley Periodicals, Inc.

PubMed ID: 28598520
Article link: Dev Dyn

Species referenced: Xenopus laevis
Genes referenced: gsc lefty lhx1 nodal1 nodal5.2 nodal5.4 pitx2 pitx3 tbxt th vegt
Antibodies: Acta1 Ab6 Pitx3 Ab1
Morpholinos: pitx3 MO1

Phenotypes: Xla Wt + pitx3 MO (Fig 10 B^1) [+]

Article Images: [+] show captions