This GFP reporter plasmid expresses Foxn4CR4.2, a noncoding sequence upstream of the Foxn4 locus.
Transcription factor forkhead box N4 (Foxn4) cooperates with key retinogenic factors to mediate the multipotent differentiation of retinal progenitors and is believed to regulate neuronal subtype diversification. FOXN4 is expressed in a subset of mitotic progenitors during retinogenesis. As such, Foxn4 controls the formation of amacrine and horizontal cells by activating the expression of the retinogenic factors MATH-3, Neuro D and PROX1. During spinal neurogenesis, the p2 progenitor domain gives rise to two intermingled distinct subtypes of interneurons, termed V2a and V2b. Foxn4 is coexpressed with the bHLH factor ASH1 (Mash1) in a subset of p2 progenitors. Functionality of Foxn4 affects ASH1 expression and regulates interneuronal formation accordingly. Over-expression of Foxn4 alone in spinal neural progenitors promotes the V2a fate at the expense of the V2b fate, whereas ASH1 suppresses both the V2a and V2b fates.
From the laboratory of Li Cai, PhD, Rutgers University.
Part of The Investigator's Annexe program.
|Gene/insert name:||Foxn4CR4.2 ? a noncoding sequence upstream of Foxn4 locus|
|Format:||Liquid/spotted on filter paper|
|Grow in E. coli at 37 C:||Yes|
|Cloning Site 5':||SpeI|
|Cloning Site 3':||FseI|
|Insert Size:||129 bp|
|Vector Backbone and Size:||basal beta-globin promoter (bGP-GFP)|
|High or low copy:||High|
CR4.2 directs GFP expression in Foxn4+ cells and differentiating horizontal cells.
Chick retinas were electroporated with either the control CAG-GFP construct or CR4.2-bGP-GFP (CR4.2-GFP) construct at embryonic day 4 (E4). Transfected retinas were harvested at E6, E7, and E8 during development, sectioned, and immunostained for GFP (green) and Foxn4 (red). Double labeled cells (boxed regions) are shown in highermagnification on the right (indicated by arrowheads; arrows point to Foxn4-negative cells). Scale bar: 20 mm.
Adapted from: Islam MM, et al. Biol Open. 2013 Sep 6;2(11):1125-36.
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