1. Establishment of HIV-1Establishment of HIV-1
resistance in T cells byresistance in T cells by
zinc-finger nucleaseszinc-finger nucleases
By: Ryla BestBy: Ryla Best

2. BackgroundBackground
 HIV-1HIV-1
 T CellsT Cells
 CCR5CCR5
 ZFNsZFNs

3. Gene TherapiesGene Therapies
 RNA inhibitorsRNA inhibitors
Antisense, ribozymes, RNAiAntisense, ribozymes, RNAi
 Protein inhibitorsProtein inhibitors
Zinc-finger, intrabodies/kinesZinc-finger, intrabodies/kines
Figure 1. (Perez, 2008)Figure 1. (Perez, 2008)

4. CCR5 MechanismCCR5 Mechanism
Figure 2. (Schoofs, 2008)Figure 2. (Schoofs, 2008)

5. Material and MethodsMaterial and Methods
 ZFNZFN
 GFP, GHOST-CCR5GFP, GHOST-CCR5
 NOG miceNOG mice
 PCRPCR
 Surveyor nuclease assaySurveyor nuclease assay
 CytometryCytometry

6. NOG Mice MethodNOG Mice Method
Figure 3. (Perez, 2008)Figure 3. (Perez, 2008)

7. Results: CCR5 ExpressionResults: CCR5 Expression
Figure 4. (Perez, 2008)Figure 4. (Perez, 2008)

8. Results: CytometryResults: Cytometry
Figure 5. (Perez, 2008)Figure 5. (Perez, 2008)

9. Results: ZFNs SurvivalResults: ZFNs Survival
Figure 6. (Perez, 2008)Figure 6. (Perez, 2008)

10. Results: NOG MiceResults: NOG Mice
Figure 7. (Perez, 2008)Figure 7. (Perez, 2008)

11. ConclusionConclusion
 ZFNs reduces CCR5 expressionZFNs reduces CCR5 expression
 Can survive during HIV-1 infectionCan survive during HIV-1 infection
 Increase in T cell countIncrease in T cell count
 Protection vs. viremiaProtection vs. viremia
(Perez, 2008)(Perez, 2008)

12. Why is it important?Why is it important?
 HIV cureHIV cure
 ImmunotherapyImmunotherapy
 Other viral curesOther viral cures

13. Questions?Questions?