1. Nobel price 2006 Andrew Z. Fire Craig C. Mello
"for their discovery of RNA interference - gene silencing by double-stranded RNA"
Andrew Z. Fire
Craig C. Mello
USA
Stanford University
School of Medicine Stanford, CA, USA
University of Massachusetts
Medical School Worcester, MA, USA

2. Serendipity?
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans Andrew Fire, SiQun Xu, Mary K. Montgomery,Steven A. Kostas, Samuel E. Driver & Craig C. Mello
NATURE | VOL 391 | 19 FEBRUARY 1998
They worked on something else in C. elegans
Gene expression – sense RNA (control)
Gene silencing – antisense RNA (weak effect)
One more controla with double strand (s-as) RNA,
SURPRISE – IT WORKED VERY WELL

3. Nobelova price experiment
dsRNA blocks target mRNA
asRNS
dsRNS

4. Not absolute new
Drosophila (Elbashir, EMBO J, 2001, Genes and Dev, 2001)
dsRNA → nt small dsRNA
small interferring RNA – siRNA
siRNS
21-25 nt dsRNA
from a bigger dsRNA precursor molecule

5. These are coded in genome – also in ours
C. elegans (Lee, 1993, Cell)
NON CODING SEQUENCES!
SMALL (61 → 22nt) RNA
lin-14 mRNS 3’ UTR komplementerita
REGULATION OF GENE EXPRESSION
microRNA (miRNA)

6. miRNS biogenesis The code is transcribed by RNA polymerase II
More possibilities
– separated - 1 miRNA
– clusters
– in INTRONS!
Mendell, 2008, Cell

7. Biogenesis and effect of siRNA
Dykxhoorn, 2003, Nat Rev Mol Cell Biol

8. Finalisation of siRNA With protein complexes (coded in genome)
Dicer makes siRNA (+ATP)
RISC – komplex with siRNA, cuts mRNA (+ATP)
Argonauta – blocks translation

9. siRNA effect
Eckstein, 2005, TIBS

10. Another possibility
Inhibícia translácie

11. miRNA - functions In human genome at least 500 – 1000 miRNa genes
1-1,5 % of all structural genes
A new (?) form of gene expression regulation – on the interface of transcription and translation
THE LAST CHECKPOINT (SMALL NINJA POLICIMEN)
miRNA expression regulation????
tissue specificity, dependent on ontogenesis
Involved in
Development, differentiation, apoptosis, cell cycle

12. miRNA functions One miRNA can regulate more mRNA (Lim, 2005, Nature)
In different conditions different picture of expression
miR-1 – heart, miR-124 – brain
In vitro experiments
miR-1 96, miR124 downregulated 174 génov

13. miRNA and disease
miRNA profiles different in normal and malignant cells
miRNA profiles in diabetes, cardiovascular and kidney diseases, degenerative disease of CNS – intensive research currently without aplication in practical medicine
SNPs in miRNA genes and disease???
Therapeutical perspectives?
Malignancies
Farmakogenetics
Design of artifical siRNA?
Not a simple task
Get them to the proper place – nanobiotechnology

14. Fewell, 2006, Drug Discovery