Identification of a novel lncRNA (G3R1) regulated by GLIS3 in pancreatic β-cells

in Journal of Molecular Endocrinology
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  • 1 Cell Biology Group, Immunity, Inflammation and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, USA
  • 2 Knockout Mouse Core, Reproductive & Developmental Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, USA

Correspondence should be addressed to A M Jetten: jetten@niehs.nih.gov
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Recent advances in high throughput RNA sequencing have revealed that, in addition to messenger RNAs (mRNAs), long non-coding RNAs (lncRNAs) play an important role in the regulation of many cell functions and of organ development. While a number of lncRNAs have been identified in pancreatic islets, their function remains largely undetermined. Here, we identify a novel long ncRNA regulated by the transcription factor GLIS3, which we refer to as GLIS3 regulated 1 (G3R1). This lncRNA was identified for its significant loss of expression in GLIS3 knockout mouse pancreatic islets. G3R1 appears to be specifically expressed in mouse pancreatic β-cells and in a β-cell line (βTC-6). ChIP-seq analysis indicated that GLIS3 and other islet-enriched transcription factors bind near the G3R1 gene, suggesting they directly regulate G3R1 transcription. Similarly, an apparent human homolog of G3R1 displays a similar expression pattern, with additional expression seen in human brain. In order to determine the function of G3R1 in mouse pancreatic β-cells, we utilized CRISPR to develop a knockout mouse where ~80% of G3R1 sequence is deleted. Phenotypic analysis of these mice did not reveal any impairment in β-cell function or glucose regulation, indicating the complexity underlying the study of lncRNA function.

Supplementary Materials

    • Supplementary Fig 1. G3R1 observed sequence differs from the annotated sequence. Sequence of G3R1, with annotated sequence marked in green and observed sequence marked in blue.
    • Supplementary Fig 2. Chronic hyperglycemia, but not acute glucose treatment, regulates G3R1. (A) Min6 cells were pre-treated with low glucose (1mM) for 1 hour, then stimulated with either low (1 mM) or high (16 mM) glucose. N=3. (B) Freshly isolated pancreatic islets from wild type (C57BL/6J) and db/db mice at 8-weeks of age of both genders. N=10 for each genotype, * indicates p<0.01. Error bars indicate standard error of the mean.
    • Supplementary Fig 3. Identification of hG3R1 conserved regions. Indicated regions of high homology on human chromosome 22. Adapted from the NCBI genome browser for hg38.
    • Supplementary Table 1. Primers and Antibodies.

 

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