1. Short hairpin RNAs
Dave Lyons
May 2009

2. Short hairpin RNAs Discovery of interfering RNAs shRNAs in biology
shRNAs in medicine

3. Short hairpin RNAs Discovery of interfering RNAs shRNAs in biology
shRNAs in medicine

4. Discovery of RNAi
. . .
1990
2000
2010

5. “Invention” of shRNA
2000
2010
Paddison et al. 2002

6. “Invention” of shRNA
Paddison et al. 2002; Rao et al. 2009

7. Short hairpin RNAs Discovery of interfering RNAs shRNAs in biology
shRNAs in medicine

8. shRNA in biological research
Large-scale screens
In vivo studies
Validation

9. shRNAs for large-scale screens
Dykxhoorn and Lieberman 2005

10. shRNAs for in vivo studies
1.) Mouse KO / KI / BAC
2.) Transplantation of infected SCs
3.) Infect mouse with vector
Dykxhoorn and Lieberman 2005

11. shRNAs for in vivo studies
4.) Tail vein injection
Pol II promoter
Pol III promoter
Dykxhoorn and Lieberman 2005; Giering et al. 2008

12. shRNA - validation 4.) Tail vein injection Off-target effects?
Pol III promoter
Pol III promoter
Pol II promoter
Pol II promoter
Dykxhoorn and Lieberman 2005; Giering et al. 2008

13. shRNA - validation
1.) need a phenotypic assay to accurately confirm on-target effect.
2.) need to control for specific and non-specific off-target effects.
- expression profiling of transfected controls
- general immune response?
- bottleneck effects (e.g. nuclear export)

14. Short hairpin RNAs Discovery of interfering RNAs shRNAs in biology
shRNAs in medicine

15. shRNA in medicine
shRNA is difficult to deliver without viruses but in vivo experiments in mouse suggest better overall for specificity than siRNA.
Successful viral targeting is central to future of shRNA therapeutics.
Clinical trials with siRNA outnumber shRNA currently.