Presentation is loading. Please wait.

Presentation is loading. Please wait.

Gfp - Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels.

Similar presentations


Presentation on theme: "Gfp - Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels."— Presentation transcript:

1 gfp - Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels

2 GFP + Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels

3 Gene correction in trans PuroeGFPPoly-lacO Cut sites trGFP nuclease PuroeGFPPoly-lacO

4 Gene correction in trans PuroeGFPPoly-lacO Cut sites trGFP nuclease PuroeGFPPoly-lacO

5 I-AniI Y2 promotes correction in trans EndoDonor% GFP + I-SceI  Ani Y   I-Sce  0.03 (75) Ani Y  0.01 (30) GFP + donor + I-SceI + donor + Ani Y2 FL2-H

6 I-AniI Y2 “nickase” promotes correction in trans I-AniIDonor% GFP + Y  Y2-nick   Y  (30) Y2-nick+0.01  (5) + donor + Ani Y2 + donor + Ani Y2-nick GFP FL2-H

7 Modulation of gene correction in trans: tethering factors at recipient PuroeGFPPoly-lacO Cut sites trGFP nuclease tethered factor tethered factor PuroeGFPPoly-lacO

8 Recipient chromatin structure regulates gene correction in trans Tethered Protein % GFP + (fold reduction) * I-SceI I-SceI + donor I-SceI + donor LacI0.20 I-SceI + donor HP1-LacI0.017 (11) * Average of two experiments + HP1-LacI GFP FL2-H + LacI

9 Modulation of gene correction in trans: tethering factors at donor PuroeGFP Cut sites trGFP nuclease tethered factor tethered factor PuroeGFP Poly-lacO

10 % HP1 tethered at donor blocks correction in trans LacO donor + ISceI LacO donor + ISceI + HP1-LacI LacO donor + ISceI + LacI % 0.22% GFP FL2 Height 40-fold decrease in recombination Jason Cummings


Download ppt "Gfp - Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels."

Similar presentations


Ads by Google