Diagenode

Transgenic mice for in vivo epigenome editing with CRISPR-based systems


Gemberling, M. et al.

The discovery, characterization, and adaptation of the RNA-guided clustered regularly interspersed short palindromic repeat (CRISPR)-Cas9 system has greatly increased the ease with which genome and epigenome editing can be performed. Fusion of chromatin-modifying domains to the nuclease-deactivated form of Cas9 (dCas9) has enabled targeted gene activation or repression in both cultured cells and in vivo in animal models. However, delivery of the large dCas9 fusion proteins to target cell types and tissues is an obstacle to widespread adoption of these tools for in vivo studies. Here we describe the generation and validation of two conditional transgenic mouse lines for targeted gene regulation, Rosa26:LSL-dCas9-p300 for gene activation and Rosa26:LSL-dCas9-KRAB for gene repression. Using the dCas9p300 and dCas9KRAB transgenic mice we demonstrate activation or repression of genes in both the brain and liver in vivo, and T cells and fibroblasts ex vivo. We show gene regulation and targeted epigenetic modification with gRNAs targeting either transcriptional start sites (TSS) or distal enhancer elements, as well as corresponding changes to downstream phenotypes. These mouse lines are convenient and valuable tools for facile, temporally controlled, and tissue-restricted epigenome editing and manipulation of gene expression in vivo.

Tags
Antibody
CRISPR

Share this article

Published
March, 2021

Source

Products used in this publication

  • CRISPR/Cas9 Antibody
    C15200229-100
    CRISPR/Cas9 Antibody
  • Bioruptor Pico
    B01080010
    Bioruptor® Pico 非接触式超声波破碎仪
  • some alt
    C01020031
    15 ml Bioruptor® Pico Tubes & sonication beads

活动

  • 64th ASH Annual Meeting and Exposition 2022
    New Orleans, Louisiana, USA
    Dec 10-Dec 13, 2022
 查看所有活动

新闻

 查看所有新闻


The European Regional Development Fund and Wallonia are investing in your future.

Extension of industrial buildings and new laboratories.


       Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy   |   Diagenode Diagnostics