1 / 16

Homology-directed gene repair

Homology-directed gene repair. exogenous repair substrate. I-SceI. xx. sister chromatids. xx. Rad51…. Sequence correction. xx. Homology-directed gene repair. exogenous repair substrate. I-SceI. xx. sister chromatids. xx. Rad51…. Sequence correction. xx. DRGFP reporter.

tareq
Download Presentation

Homology-directed gene repair

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. Homology-directed gene repair exogenous repair substrate I-SceI xx sister chromatids xx Rad51… Sequence correction xx

  2. Homology-directed gene repair exogenous repair substrate I-SceI xx sister chromatids xx Rad51… Sequence correction xx

  3. DRGFP reporter CA puroR I-SceI STOP BcgI gene conversion CA GFP+ puroR BcgI BcgI

  4. 041408 293T stable DRGFP (S8) pCMV-I-SceI no DNA 48 hrs 72 hrs

  5. Direct Repeat Reporters:recombination in cis

  6. 041408 293T DRGFP (cis) DRGFP+I-SceI I-SceI DRGFP

  7. 041408 293T DRGFP-lacO (cis) DRGFPlacO+I-SceI DRGFPlacO

  8. Exogenous Donor Reporter:recombination in trans

  9. 041408 293T ZFGFP + iGFP (trans) ZFGFP+iGFP ZFGFP+iGFP+I-SceI

  10. 041408 293T ZFGFP-lacO + iGFP (trans) ZFGFP-lacO ZFGFP-lacO+iGFP+I-SceI

  11. Too Much DNA?

  12. 050208 293T: Efficiency of Transient Transfection no DNA 62 ng eGFP 125 ng eGFP 250 ng eGFP 500 ng eGFP 1.0 g eGFP 2.0 g eGFP 4.0 g eGFP

  13. “Stable” DRGFP?

  14. “Stable” DRGFP (puroS) 48 hrs 73 hrs 98 hrs No DNA CMV-Sce

  15. “Stable” DRGFP (puroR) 48 hrs 73 hrs 98 hrs No DNA CMV-Sce

  16. Plans • Lines with integrated trans reporter • Enhance repair by tethering chromatin modifiers and repair factors • Back to B-cells

More Related