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JoVE Journal
Bioengineering

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In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
 

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Article DOI: 10.3791/64403-v 10:44 min May 5th, 2023
May 5th, 2023

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Summary

Here, we present a protocol for the in vitro selection of engineered transcriptional repressors (ETRs) with high, long-term, stable, on-target silencing efficiency and low genome-wide, off-target activity. This workflow allows for reducing an initial, complex repertoire of candidate ETRs to a short list, suitable for further evaluation in therapeutically relevant settings.

Tags

Bioengineering Targeted Epigenetic Silencing Gene Inactivation RNA Interference Artificial Nucleases DNA Double Strand Break Epigenetic Editing Programmable DNA-binding Domain KRAB Domain DNMT3A DNMT3L Heritable Repressive Epigenetic States Hit-and-run Nature DNA Demethylation
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