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Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins.

Rolen M Quadros | Hiromi Miura | Donald W Harms | Hisako Akatsuka | Takehito Sato | Tomomi Aida | Ronald Redder | Guy P Richardson | Yutaka Inagaki | Daisuke Sakai | Shannon M Buckley | Parthasarathy Seshacharyulu | Surinder K Batra | Mark A Behlke | Sarah A Zeiner | Ashley M Jacobi | Yayoi Izu | Wallace B Thoreson | Lisa D Urness | Suzanne L Mansour | Masato Ohtsuka | Channabasavaiah B Gurumurthy
Genome biology | 2017

Conditional knockout mice and transgenic mice expressing recombinases, reporters, and inducible transcriptional activators are key for many genetic studies and comprise over 90% of mouse models created. Conditional knockout mice are generated using labor-intensive methods of homologous recombination in embryonic stem cells and are available for only ~25% of all mouse genes. Transgenic mice generated by random genomic insertion approaches pose problems of unreliable expression, and thus there is a need for targeted-insertion models. Although CRISPR-based strategies were reported to create conditional and targeted-insertion alleles via one-step delivery of targeting components directly to zygotes, these strategies are quite inefficient.

Pubmed ID: 28511701

Associated grants

  • Agency: NCI NIH HHS, United States
    Id: P30 CA036727
  • Agency: NIGMS NIH HHS, United States
    Id: P20 GM103471
  • Agency: NEI NIH HHS, United States
    Id: R01 EY010542
  • Agency: Wellcome Trust, United Kingdom
    Id: 087377
  • Agency: NIGMS NIH HHS, United States
    Id: P30 GM110768
  • Agency: NIDCD NIH HHS, United States
    Id: R21 DC014779

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


Ambion Inc. (tool)

RRID:SCR_008406

A division of Applied Biosystems selling products for the isolation, detection, quantification, amplification, and characterization of RNA.

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Adobe Photoshop (tool)

RRID:SCR_014199

Software for image processing, analysis, and editing. The software includes features such as touch capabilities, a customizable toolbar, 2D and 3D image merging, and Cloud access and options.

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CHOPCHOP (tool)

RRID:SCR_015723

Web application for designing gRNAs for CRISPR/Cas9 experiments. It selects target sites for CRISPR/Cas9, CRISPR/Cpf1 or TALEN-directed mutagenesis.

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International Mouse Phenotyping Consortium (IMPC) (tool)

RRID:SCR_006158

Center that produces knockout mice and carries out high-throughput phenotyping of each line in order to determine function of every gene in mouse genome. These mice will be preserved in repositories and made available to scientific community representing valuable resource for basic scientific research as well as generating new models for human diseases.

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C57BL/6J (tool)

RRID:IMSR_JAX:000664

Mus musculus with name C57BL/6J from IMSR.

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B6.Cg-Gt(ROSA)26Sortm1.3(CAG-tdTomato.-EGFP)Pjen/J (organism)

RRID:IMSR_JAX:026932

Mus musculus with name B6.Cg-Gt(ROSA)26Sortm1.3(CAG-tdTomato.-EGFP)Pjen/J from IMSR.

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