CCR5 Zincfinger

Establishment of HIV-1 Establishment of HIV-1 resistance in T cells by resistance in T cells by zinc-finger nucleaseszinc-finger nucleases

By: Ryla BestBy: Ryla Best

BackgroundBackgroundHIV-1HIV-1T CellsT CellsCCR5CCR5ZFNsZFNs

Gene TherapiesGene TherapiesRNA inhibitorsRNA inhibitors

Antisense, ribozymes, RNAiAntisense, ribozymes, RNAiProtein inhibitorsProtein inhibitors

Zinc-finger, intrabodies/kinesZinc-finger, intrabodies/kines

Figure 1. (Perez, 2008)Figure 1. (Perez, 2008)

CCR5 MechanismCCR5 Mechanism

Figure 2. (Schoofs, 2008)Figure 2. (Schoofs, 2008)

Material and MethodsMaterial and MethodsZFNZFNGFP, GHOST-CCR5GFP, GHOST-CCR5NOG miceNOG mice

PCRPCRSurveyor nuclease assaySurveyor nuclease assayCytometryCytometry

NOG Mice MethodNOG Mice Method

Figure 3. (Perez, 2008)Figure 3. (Perez, 2008)

Results: CCR5 ExpressionResults: CCR5 Expression

Figure 4. (Perez, 2008)Figure 4. (Perez, 2008)

Results: CytometryResults: Cytometry

Figure 5. (Perez, 2008)Figure 5. (Perez, 2008)

Results: ZFNs SurvivalResults: ZFNs Survival

Figure 6. (Perez, 2008)Figure 6. (Perez, 2008)

Results: NOG MiceResults: NOG Mice

Figure 7. (Perez, 2008)Figure 7. (Perez, 2008)

ConclusionConclusionZFNs reduces CCR5 expressionZFNs reduces CCR5 expressionCan survive during HIV-1 infectionCan survive during HIV-1 infection Increase in T cell countIncrease in T cell countProtection vs. viremiaProtection vs. viremia

(Perez, 2008)(Perez, 2008)

Why is it important?Why is it important?HIV cureHIV cure ImmunotherapyImmunotherapyOther viral curesOther viral cures

Questions?Questions?