Human Genetics And Molecular Medicine - Master Dissertation
Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/104
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Item Potential Mitochondrial-Specific Function Of piRNAs(Central University of Punjab, 2018) Paul, Shouvik; Singh, SandeepPiwi-interacting RNAs (piRNAs) are (26-31 nt) small noncoding RNAs processed from their longer precursor transcripts with the help of Piwi proteins. There are more than 30,000 piRNA genes present in the human genome which now turns out to be emerging player in both homeostasis and diseases. Localization of piRNA and PIWI in the repeat region of the mammalian nuclear genome in germ cells has been reported, although localization and potential functional role of piRNA in the mammalian mitochondrial genome are largely unknown. We have taken 111 piRNA sequences found in the MCF-7 mitochondrial genome, which is obtained by NGS analysis for alignment study. Resulting piRNA have been aligned with DQ112870 North American Homo sapiens mitochondrion genome for studying post- transcriptional roles of piRNA.Item Generation of Rho-0 Cells using MDA-MB-231 Cell Line and Measurement of Drug Cytotoxicity(Central University of Punjab, 2018) Sharma, Bharti; Singh,SandeepThe ATP generation via Oxidative phosphorylation (OXPHOS) system located in the inner membrane of mitochondria, is regulated by the coordinated interaction between nucleus and mitochondria. In the same context, mitochondrial-depleted cell (Rho-0) can be a helpful approach to study the mitochondrial metabolism, mitochondrial role in various cellular processes such as apoptosis, mitochondrial role in various mitochondrial related disorders and cancer. To generate Rho-0 cells, EtBr mediated mtDNA depletion was done and verified by agarose gel electrophoresis. % cell viability, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) production was measured after 24 hr treatment with 3 drugs, ?-amanitin, Doxorubicin and DCA in both parental MDA-MB-231 and Rho-0 cells. Reduced cell death and ROS production was observed in Rho-0 cells indicating the resistance against apoptosis in Rho-0 cells and demonstrating the possible role of mitochondria in intrinsic pathway of apoptosis. MMP was observed to be maintained in Rho-0 cells indicating the role of nuclear genome in the maintenance of MMP.