In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

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1.3K Views

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10:44 min

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May 5th, 2023

DOI :

10.3791/64403-v

May 5th, 2023

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Alessandro Migliara¹, Martino Alfredo Cappelluti¹, Francesca Giannese², Sara Valsoni¹, Alberto Coglot¹, Ivan Merelli¹^,³, Davide Cittaro², Angelo Lombardo¹^,⁴

¹San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute, ²Center for Omics Sciences, IRCCS San Raffaele Institute, ³Institute for Biomedical Technologies, National Research Council, ⁴Vita-Salute San Raffaele University

Podsumowanie

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Epigenetic Silencing

Gene Silencing Technologies

Engineered Transcriptional Repressors

RNA Interference

Gene Disruption

Artificial Nucleases

Double strand DNA Break

Chromatin Compaction

Transcriptional Inhibition

CRISPR Cas9

Fluorophore Expression Cassette

K 562 Cell Line

Rozdziały w tym wideo

0:05

Introduction

1:34

Engineering a Fluorescence-Based Reporter Cell Line

3:10

Designing gRNAs for CRISPR/dCas9-Based Epigenetic Silencing of Target Gene

4:13

Arrayed Transient Delivery of CRISPR/dCas9-Based ETRs in Reporter Cell Line

7:19

Analyzing Transcriptional Activity of Target Gene Over Time

8:11

Results: dCas9 ETR-Mediated Epigenetic Silencing of Human B2M Gene by gRNAs

9:23

Conclusion

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