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Dev Dyn January 1, 2017; 246 (9): 657-669.

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 .

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. 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. 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, 2017. © 2017 Wiley Periodicals, Inc.

PubMed ID: 28598520
Article link: Dev Dyn

Genes referenced: gsc lefty lhx1 nodal1 nodal5 nodal5.2 pitx2 pitx3 tbxt th vegt

Antibodies referenced: Acta1 Ab6 Pitx3 Ab1
Morpholinos referenced: pitx3 MO1

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