| First Author | Alzhanov D | Year | 2016 |
| Journal | Physiol Genomics | Volume | 48 |
| Issue | 2 | Pages | 167-72 |
| PubMed ID | 26645089 | Mgi Jnum | J:230726 |
| Mgi Id | MGI:5763683 | Doi | 10.1152/physiolgenomics.00095.2015 |
| Citation | Alzhanov D, et al. (2016) Characterizing a distal muscle enhancer in the mouse Igf2 locus. Physiol Genomics 48(2):167-72 |
| abstractText | Insulin-like growth factor-2 (IGF2) is highly expressed in skeletal muscle and was identified as a quantitative trait locus for muscle mass. Yet little is known about mechanisms of its regulation in muscle. Recently, a DNA segment found approximately 100 kb from the Igf2 gene was identified as a possible muscle transcriptional control element. Here we have developed an in vivo reporter system to assess this putative enhancer by substituting nuclear (n) EGFP for Igf2 coding exons in a bacterial artificial chromosome containing the mouse Igf2 - H19 chromosomal locus. After stable transfection into a mesenchymal stem cell line, individual clones were converted to myoblasts and underwent progressive muscle-specific gene expression and myotube formation in differentiation medium. Transgenic mRNA and nuclear-targeted enhanced green fluorescent protein were produced coincident with endogenous Igf2 mRNA, but only in lines containing an intact distal conserved DNA element. Our results show that a 294 bp DNA fragment containing two E-boxes is a necessary and sufficient long-range enhancer for induction of Igf2 gene transcription during skeletal muscle differentiation and provides a robust experimental platform for its further functional dissection. |
