Biochemical analysis of engineered TAL effector nuclease at on and novel off target sites
Abstract/Contents
- Abstract
- Genome editing is a process that entails precise manipulation of a DNA sequence. To achieve this, a define stretch of nucleotides is replaced by a new, exogenously provided, DNA template using the cellular DNA repair machinery as a mean to promote the exchange. By taking this approach, we not only can repair alterations that impede gene functions but we can also expand our knowledge of basic biological processes driven by genes. Different classes of site-specific hybrid nucleases have been engineered precisely for the purpose of deleting a DNA sequence in a targeted manner. There are three main supporting platforms, Zinc Finger Nucleases (ZFNs), Transcription Activator Like Effector Nucleases (TALENs) and Cluster Regulatory Interspaced Short Palindromic Repeats/Cas9 Nucleases (CRISPR/Cas9). Of these three, TALENs have the highest targeting range, the flexibility of modifying different genomes and cell types with the lowest off-target activity, features that promote TALENs as potential powerful therapeutic tools. To further characterize them, I have employed a biochemical approach that allowed me to determine their specificity, affinities and kinetics at both cognate and novel OFF target DNA binding sites. From this study it became apparent that TALENs' kinetic signature is the missing link that connects their in vitro behavior with in vivo activity.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2015 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Pavel-Dinu, Mara |
|---|---|
| Associated with | Stanford University, Department of Biology. |
| Primary advisor | Porteus, Matthew H |
| Thesis advisor | Porteus, Matthew H |
| Thesis advisor | Gozani, Or Pinchas |
| Thesis advisor | Greenleaf, William James |
| Thesis advisor | Mudgett, Mary Beth, 1967- |
| Advisor | Gozani, Or Pinchas |
| Advisor | Greenleaf, William James |
| Advisor | Mudgett, Mary Beth, 1967- |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Mara Pavel-Dinu. |
|---|---|
| Note | Submitted to the Department of Biology. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2015. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2015 by Mara Pavel-Dinu
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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