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. 2011 Aug 8;39(20):e133. doi: 10.1093/nar/gkr628

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — RNA polymerase II-driven plasmids can be used to express a single transcript encoding both a fluorescent protein and an effective artificial miRNA in chicken embryos. (A) Schematic of the plasmid constructs used in this study. The stem–loop structure of the miRNA30-like cassette is shown for an artificial miRNA against Shh (miShh). The target (sense) sequence of Shh is indicated in red. (B) Immunolabeling of cryosections of untreated chicken spinal cords at HH26 with the 5E1 antibody shows normal distribution of Shh in the notochord (nc), floor plate (asterisks) and adjacent ventral spinal cord. The boxed area is shown in higher power in (C), and equivalent regions are shown in (D–G). (D and F) RFP-positive cells in the floor plate and ventral spinal cord of embryos bilaterally electroporated with the control βactin-RFP construct (D) or βactin-RFP-miShh (E). The corresponding images showing Shh expression in each condition are shown in (F) and (G) (compare to C). Scale bar: 100 µm.