Poster abstracts

Poster number 65 submitted by Chaucola Pleasant

Splicing Interference (SPLICEi) By Small Nuclear RNAs in Breast Cancer

Chaucola K. Pleasant (BGES, Cleveland State University), Girish C. Shukla (BGES, Cleveland State University)

Abstract:
In nuclear pre-mRNA splicing, introns are removed through the work of small nuclear RNAs (snRNAs). In the human genome there are two types of introns. Major class introns, the U2-dependent type, which are spliced by U1, U2, U4, U5 and U6 snRNAs, and the minor class introns, the U12 dependent type, which are spliced by U11, U12, U4atac, U5, and U6atac snRNAs. It has been observed that over expression of minor class spliceosomal snRNAs can have an inhibitory effect on the splicing of major class introns. To further test the concept we targeted the Human Epidermal Receptor 2(HER-2)/Neu proto-oncogene, which is over- expressed in 20-30% of breast tumors. We constructed a series of mutant human U6atac and U11 snRNAs to target introns 1, 2, 6, 8, 12 and 13 of HER-2/Neu to block nuclear pre-mRNA splicing and down regulate protein synthesis. We transfected mutant snRNAs in breast cancer cell line MDA-MB-453 to determine the efficacy of our approach on HER-2/Neu pre-mRNA splicing interference. Our preliminary data indicates variable effect on the HER-2/Neu pre-mRNA splicing and protein synthesis in cultured cells. Further experiments are in progress to confirm the efficacy of mutant snRNAs in blockading of HER-2/Neu expression.

Keywords: snRNA, splicing, Breast Cancer