Syllabus |
The contents below will be applied in the realization of a mini-project that aims to use a spermatogonial cell line (GC-spg1) to evaluate the effect of siRNA-mediated knockdown of estrogen receptor on the expression of c-kit, a gene associated with cell proliferation. 1. Regulation of gene expression: promoters, enhancers/silencers, insulators, mediators, coregulators, mechanisms of alternative splicing, interference RNA, miRNAs, lncRNAs, chromatin remodeling, and CpG methylation. 2. Advanced tools in DNA editing to control gene expression: siRNAs, shRNAs, and CRISPR. 3. Sample preparation for nucleic acid and protein extraction. 4. Real-time PCR: fundamentals, detection methods, primer design, analysis of primer specificity and efficiency, normalization, absolute and relative quantification, result analysis. 5. Western blot: fundamentals, detection methods, antibody selection, normalization, relative quantification, and result analysis.
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