Optimizations of Type 1 Collagen RNA in vitro Transcriptions
Additional Funding Sources
The project described was supported by the Center of Excellence in Biomedical Research through the Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant Nos. P20GM109095 and P20GM103408 and the National Science Foundation S-STEM Gateway Scholarships in Biological Sciences under Grant Award No. DUE-1644233.
Presentation Date
7-2022
Abstract
Current studies on the defense against chronic fibrous have focused on the inhibition of the inflammatory pathways. However, it is believed that an alternative treatment method could be found by inhibiting the overexpression of collagen. A new defense pathway could be discovered by analyzing collagen production and its relationship with a proposed chaperone protein, LARP6. The production of the collagen 1α1(I), 1α2(I), 3α1(I), and 5α2(I) proteins is facilitated by RNA strains containing a highly conserved 5' Stem Loop (5'SL) in the untranslated region. It is believed that LARP6 helps assist in folding the RNA into the stem-loop conformation at which point protein synthesis can initiate. To better understand the effect of LARP6 on collagen production, a deeper understanding of the RNA 5'SL’s 3D structure must be determined. However, prior to analysis, a large stock of purified RNA must be transcribed and isolated. This research project tested a series of collagen RNA transcription reactions under various constraints. By optimizing the transcription conditions, large-scale reactions of collagen RNA can be performed with a high yield. After a large volume of RNA is transcribed, the 5' SL analysis can proceed by nuclear magnetic resonance (NMR).
Optimizations of Type 1 Collagen RNA in vitro Transcriptions
Current studies on the defense against chronic fibrous have focused on the inhibition of the inflammatory pathways. However, it is believed that an alternative treatment method could be found by inhibiting the overexpression of collagen. A new defense pathway could be discovered by analyzing collagen production and its relationship with a proposed chaperone protein, LARP6. The production of the collagen 1α1(I), 1α2(I), 3α1(I), and 5α2(I) proteins is facilitated by RNA strains containing a highly conserved 5' Stem Loop (5'SL) in the untranslated region. It is believed that LARP6 helps assist in folding the RNA into the stem-loop conformation at which point protein synthesis can initiate. To better understand the effect of LARP6 on collagen production, a deeper understanding of the RNA 5'SL’s 3D structure must be determined. However, prior to analysis, a large stock of purified RNA must be transcribed and isolated. This research project tested a series of collagen RNA transcription reactions under various constraints. By optimizing the transcription conditions, large-scale reactions of collagen RNA can be performed with a high yield. After a large volume of RNA is transcribed, the 5' SL analysis can proceed by nuclear magnetic resonance (NMR).