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 PMID:25690852  

Inducible in vivo genome editing with CRISPR-Cas9.

Lukas E Dow | Jonathan Fisher | Kevin P O'Rourke | Ashlesha Muley | Edward R Kastenhuber | Geulah Livshits | Darjus F Tschaharganeh | Nicholas D Socci | Scott W Lowe
Nature biotechnology | 2015

CRISPR-Cas9-based genome editing enables the rapid genetic manipulation of any genomic locus without the need for gene targeting by homologous recombination. Here we describe a conditional transgenic approach that allows temporal control of CRISPR-Cas9 activity for inducible genome editing in adult mice. We show that doxycycline-regulated Cas9 induction enables widespread gene disruption in multiple tissues and that limiting the duration of Cas9 expression or using a Cas9(D10A) (Cas9n) variant can regulate the frequency and size of target gene modifications, respectively. Further, we show that this inducible CRISPR (iCRISPR) system can be used effectively to create biallelic mutation in multiple target loci and, thus, provides a flexible and fast platform to study loss-of-function phenotypes in vivo.

Pubmed ID: 25690852

Research resources used in this publication

None found

Additional research tools detected in this publication

Antibodies used in this publication

None found

Associated grants

  • Agency: NIGMS NIH HHS, United States
    Id: T32GM07739
  • Agency: NCI NIH HHS, United States
    Id: F31 CA192835
  • Agency: NCI NIH HHS, United States
    Id: F32 CA177072
  • Agency: NCI NIH HHS, United States
    Id: P30 CA008748
  • Agency: NCI NIH HHS, United States
    Id: P01 CA087497
  • Agency: NCI NIH HHS, United States
    Id: 1K22CA181280-01
  • Agency: NCI NIH HHS, United States
    Id: P01 CA013106
  • Agency: NIGMS NIH HHS, United States
    Id: T32 GM007739
  • Agency: NCI NIH HHS, United States
    Id: R01 CA195787
  • Agency: NCI NIH HHS, United States
    Id: P01 CA129243
  • Agency: NCI NIH HHS, United States
    Id: 1F32CA177072-01
  • Agency: NCI NIH HHS, United States
    Id: CA-013106
  • Agency: Howard Hughes Medical Institute, United States

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This is a list of tools and resources that we have found mentioned in this publication.


SHRiMP (tool)

RRID:SCR_005496

A software package for aligning genomic reads against a target genome.

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Dako (tool)

RRID:SCR_013530

An Antibody supplier; Dako was purchased by Agilent in 2012 and several years later the websites began to reflect the Dako products as part of the Agilent catalog.

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Illumina MiSeq System (tool)

RRID:SCR_016379

MiSeq System offers wide range of sequencing applications. Capable of automated paired end reads and up to 15 Gb per run, delivering over 600 bases of sequence data per read. The library prep kits are optimized for variety of applications, including targeted gene, small genome, and amplicon sequencing, 16S metagenomics.

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