| Chapter title |
Purified Protein Delivery to Activate an Epigenetically Silenced Allele in Mouse Brain
|
|---|---|
| Chapter number | 12 |
| Book title |
Epigenome Editing
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7774-1_12 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7773-4, 978-1-4939-7774-1
|
| Authors |
Benjamin Pyles, Barbara J. Bailus, Henriette O’Geen, David J. Segal |
| Abstract |
The ability to activate or repress specific genes in the brain could have a tremendous impact for understanding and treating neurological disorders. Artificial transcription factors based on zinc finger, TALE, and CRISPR/Cas9 programmable DNA-binding platforms have been widely used to regulate the expression of specific genes in cultured cells, but their delivery into the brain represents a critical challenge to apply such tools in live animals. In previous work, we developed a purified, zinc finger-based artificial transcription factor that could be injected systemically, cross the blood-brain barrier, and alter expression of a specific gene in the brain of an adult mouse model of Angelman syndrome. Importantly, our mode of delivery produced widespread distribution throughout the brain. Here we describe our most current methods for the production and purification of the factor, dosage optimization, and use of live animal fluorescence imaging to visualize the kinetics of distribution. |
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