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Rapid RNA‐Protein Interaction Detection with RNA5.0 of HCMV
Author(s) -
Mickens Kassandra,
Mickens Kaylee,
Kulesza Caroline
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.778.7
Subject(s) - rna , biotinylation , biology , computational biology , virology , fusion protein , rna binding protein , microbiology and biotechnology , genetics , recombinant dna , gene
CMV is the most common cause of both congenital infection and post‐transplant infection worldwide. CMV infection can cause severe symptoms: pneumonia, encephalitis, and inflammation of vital organs. There is currently no vaccine for CMV. An understanding of how CMV progresses from its acute phase to persistence would illuminate features of the pathogen that could lead to novel treatments for the disease which block the virus from reaching latency . RNA5.0 is a lncRNA produced during CMV infection. Studies using the homologous RNA7.2 a mouse model has shown when the RNA is not expressed in vivo , CMV infection cannot reach its persistent phase. However, how it performs this function remains unclear. Our study hopes to help the field to move forward by expanding the limited knowledge about RNA5.0's function in viral persistence at a molecular level by purifying and identifying the proteins . We propose identifying proteins that bind to RNA5.0 with known cellular functions will allow us to narrow the speculation surrounding RNA5.0's function, as their functions are likely related. To purify these proteins, we will utilize the RaPID (RNA–protein interaction detection) method outlined in Ramanathan et al . 2018. In this novel approach, RNA stem loops flank RNA sequence of interest and recruits the RaPID fusion protein that biotinylates proteins proximal to inserted RNA. A streptavidin pulldown is used to capture biotinylated proteins for analysis by mass spectrometry (MS). Because the RaPID method has limited reach, in order to characterize all the proteins that bind to the 5kb lncRNA of CMV we will use 50 overlapping 150bp segments with 50bp overlaps. Support or Funding Information Fort Lewis College Undergraduate Research, Scholarship, and Creative Activity Grant This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .