Evaluation of miR-302 promoter activity in transgenic mice and pluripotent stem cell lines
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Evaluation of miR-302 promoter activity in transgenic mice and pluripotent stem cell lines Karim Rahimi 1 & Sara Parsa 2 & Mehrnoush Nikzaban 3 & Behnoush Khaledian 4 & Seyed Javad Mowla 2 & Fardin Fathi 4 Received: 21 June 2019 / Accepted: 29 September 2020 / Published online: 18 November 2020 / Editor: Tetsuji Okamoto # The Society for In Vitro Biology 2020
Abstract Some miRNAs, including the miR-302 cluster, are critical regulators of the stemness state of embryonic stem cells and cell fate patterning. In this study, we evaluated the activity of the miR-302 core promotor in mice and human pluripotent stem cells, somatic tissue derivatives, and generated transgenic mice expressing EGFP under a miR-302 promoter. The expression of EGFP under the control of the miR-302 promotor was examined in the cell lines and somatic tissues of transgenic mice, transgenic blastocysts, and embryonic stem cells derived from transgenic blastocysts. Our results showed that the miR-302 promoter is highly expressed in the mouse and human pluripotent cells, weakly expressed in the somatic tissue derivatives, is highly expressed in both blastocysts and the first passages of transgenic embryonic stem cells, and lowly expressed in the somatic tissues of transgenic mice. It can be concluded that different temporal and spatial gene expression patterns occur during the embryonic and adult stages of cells in mice. Keywords Embryonic stem cells . Adult stem cells . Transgenic mice . MicroRNAs . Blastocyst
Introduction MicroRNAs (miRNAs) are short, noncoding RNAs that execute posttranscriptional repression by directly annealing to the complementary sequences of their target RNA (Lewis et al. 2005). They are considered vital for development and the regulation of cell proliferation, are associated with disease (Wang et al. 2007), and are increasingly recognized as potential stem cell markers (Song et al. 2017; Lou et al. 2018). The miR-302a-d/367 cluster (miR-302s) is transcribed from a Pol II promoter. The primary RNA consists of 2–3 exons in humans and two exons in mice. MiR-302s is a polycistronic miRNA cluster that includes miR-302b/c/a/d and miR-367,
which derive from the same noncoding host transcript and share the same promoter and expression regulatory elements (Barroso-delJesus et al. 2008; Rahimi et al. 2018). All five miRNAs are derived from an intronic sequence. MiR-302a–d are highly related and share the same seed sequence, and this is also shared with miR-290-295 in mice and miR-373 in humans. MiR-367 is derived from the same intron sequence (Rosa and Brivanlou 2009); however, its seed sequence is not the same as the other miRNAs in the cluster and has different targets in the transcriptome. MiR-367 regulates Smad7 and TGF-β signaling in human pancreatic cancer cells, which promotes invasion and the metastasis of these cancer cells (Zhu et al. 2015). Also, miR-367 induces proliferation and stem
* Fardin Fathi [email protected] Karim Rahimi [email protected] Sara Parsa [email protected]
Seyed Javad Mowla [email protected] 1
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