SUBJECT

Title

RNA interference

Type of instruction

lecture

Level

master

Faculty

Faculty of Science

Part of degree program

Biology MSc

Credits

Recommended in

Semesters 1-4

Typically offered in

Autumn/Spring semester

Course description

I1. Historical ground (Nobel prize in Medicine in 2006). Recovery, Co-suppression, Quelling, PTGS (post transcriptional gene silencing), sense RNA, antisense RNA, double-stranded RNA. siRNA, miRNA, molecular mechanisms, similarities and differences. Applications of gene silencing in genetic analysis (phenocopy), epistasis, analysis of genetic pathways

2. Molecular mechanism of RNA interference I: dsRNA, Dicer, siRNA, RISC.

3. Molecular mechanism of RNA interference II: transcriptional and posttranscriptional gene “silencing”, RNA degradation.

4. Molecular mechanism of RNA interference III: DNA mutilation and histone modification.

5. Molecular mechanism of RNA interference IV: systemic RNA interference, transient RNA interference, inherited RNA interference.

6. Evolutionary aspects of RNA interference, conserved regulatory elements, roles in defending against viral infection, viral surviving strategies. Maintenance of genome stability, transposon silencing, retrotransposons and retroviruses.

7. Practical applications of gene silencing: RNA interference in C. elegans, D. melanogaster and mammals. Genome-wide RNA interference (knockdown) screens. Inherited gene silencing.

8. miRNAs in genomes, prediction and validation. Developmental roles for miRNAs in nematodes, insects, plants and mammals. Regulation of heterochronic genes.

9. Quantification of miRNAs (Northern analysis, RT-PCR). Functional analysis of miRNAs.

10. RNA interference in nematodes (C. elegans), insects (D. melanogaster), plants (A. thaliana), mice (M. musculus) and human cell cultures.

11. Medical applications of RNAi I. Tumour therapies. Defence against viral infections. Utilization in HIV infection.

12. Medical applications of RNAi II. Genetic analysis of inherited human diseases.

13. Practical applications of RNA interference. Utilization of RNAi in pest control. RNAi against viral infection in plants. Storable tomato. Genetic engineering of agricultural plants with elevated stress resistance.

14. Viral suppressors of RNA „silencing”. The p19 protein.

Readings

The material of the lectures
Patrick J. Paddison, Peter K. Vogt: RNA Interference, Springer Science & Business Media, 2008, ISBN 9783540751571