1 / 5

Progress Report on Cleavage Specificities of I-CreI and I-MsoI Libraries

This report presents the findings on in vivo and in vitro cleavage specificities of I-CreI and I-MsoI libraries. It includes the construction of degenerate site libraries, single bp mismatch matrix for I-MsoI, and generation of Mso mutants with higher activity. The study also delves into rational design and bacterial selection in determining the cleavage specificity of mMsoI. Key insights on mutations and target sites are discussed in detail.

keren
Télécharger la présentation

Progress Report on Cleavage Specificities of I-CreI and I-MsoI Libraries

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. NGEC Progress ReportHui Li, Monnat Group • In vivo and in vitro cleavage specificities • Construction of degenerate site library for I-CreI and I-MsoI I-CreI library: ~ 107 unique sites; 6.8 ± 2.5 mutations/per site I-MsoI library: ~ 5 ×106 unique sites; 8.2 ± 2.1 mutations/per site • Construction of single bp mismatch matrix for I-MsoI In vivo cleavage specificity based on recombination In vitro cleavage specificity • Generation of Mso mutants with higher activity • rational design • bacterial selection

  2. In vivo cleavage specificity of mMsoI

  3. In vitro cleavage specificity of Mso-I Each target site 7.5 nm

  4. In vitro cleavage specificity of mMsoI Each target site 7.5 nm

  5. Rational design of I-MsoI Mutants D81 D146, E147 E147 D146 J. M. B. (2003) 329, 253-269

More Related