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CTNNA1 is Expressed in U937 Cells and is Regulated by miR9
Author(s) -
Patel Nikhil,
Rabquer Bradley James
Publication year - 2018
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.2018.32.1_supplement.752.9
Subject(s) - untranslated region , gene , gene expression , biology , microbiology and biotechnology , messenger rna , genetics
Rheumatoid arthritis (RA), a chronic, auto‐immune, inflammatory disorder characterized by progressive joint destruction, affects 0.3–1% of people worldwide and is a major cause of disabilities. Monocyte (MN) trafficking into inflamed joints is critical to the pathogenesis of RA. Micro RNAs (miRNAs) are approximately 22‐nucleotide long single‐stranded, noncoding RNA which bind to mRNA, thus making them act as gene expression regulators. miRNAs bind to complementary sites on the 3′‐end of target mRNAs and either block their translation or enhance their degradation, resulting in downregulation of certain genes. Previous studies in our lab have identified a gene of interest called CTNNA1, as prior research has suggested miR9 to be a suitable binding match. CTNNA1 plays a role in several pathological processes, including muscle contraction and tumor metastasis. It's hypothesized that miR9 binds to CTNNA1 and that miR9 will mediate its degradation. Methods and Results U937 monocytes were cultured, mRNA was isolated, and cDNA was prepared. During this, the DNA sequence of CTNNA1 in humans was analyzed so primers could be designed to bind to the predicted 3′ UTR binding sites of miR9. PCR was then performed to yield the isolated, predicted binding site, and then base pair size was confirmed via electrophoresis gels. The gene product was then cut with restriction enzymes Sac I and Sal I, ligated into the pmirGLO vector, and was transformed into E. coli cells and plated for incubation. Selected E. coli colonies expressing the gene were grown and incubated in overnight cultures. Conclusion CTNNA1 is expressed by U937 cells and it's 3′ UTR has been cloned into pmirGLO for mir9 binding assessment. Support or Funding Information Albion College Biology Department and the Foundation for Undergraduate Research, Scholarly, and Creative Activity This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .