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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

Overview of attention for article published in Genome Biology (Online Edition), May 2017
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About this Attention Score

  • In the top 5% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (97th percentile)

Mentioned by

news
6 news outlets
blogs
4 blogs
twitter
108 tweeters
facebook
2 Facebook pages
wikipedia
1 Wikipedia page
googleplus
1 Google+ user

Citations

dimensions_citation
282 Dimensions

Readers on

mendeley
639 Mendeley
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Title
Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins
Published in
Genome Biology (Online Edition), May 2017
DOI 10.1186/s13059-017-1220-4
Pubmed ID
Authors

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

Abstract

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. Here we describe Easi-CRISPR (Efficient additions with ssDNA inserts-CRISPR), a targeting strategy in which long single-stranded DNA donors are injected with pre-assembled crRNA + tracrRNA + Cas9 ribonucleoprotein (ctRNP) complexes into mouse zygotes. We show for over a dozen loci that Easi-CRISPR generates correctly targeted conditional and insertion alleles in 8.5-100% of the resulting live offspring. Easi-CRISPR solves the major problem of animal genome engineering, namely the inefficiency of targeted DNA cassette insertion. The approach is robust, succeeding for all tested loci. It is versatile, generating both conditional and targeted insertion alleles. Finally, it is highly efficient, as treating an average of only 50 zygotes is sufficient to produce a correctly targeted allele in up to 100% of live offspring. Thus, Easi-CRISPR offers a comprehensive means of building large-scale Cre-LoxP animal resources.

Twitter Demographics

The data shown below were collected from the profiles of 108 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 639 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Netherlands 1 <1%
Germany 1 <1%
Switzerland 1 <1%
Brazil 1 <1%
Unknown 635 99%

Demographic breakdown

Readers by professional status Count As %
Researcher 149 23%
Student > Ph. D. Student 132 21%
Student > Bachelor 53 8%
Student > Master 46 7%
Other 34 5%
Other 98 15%
Unknown 127 20%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 210 33%
Agricultural and Biological Sciences 157 25%
Neuroscience 42 7%
Medicine and Dentistry 30 5%
Immunology and Microbiology 22 3%
Other 46 7%
Unknown 132 21%

Attention Score in Context

This research output has an Altmetric Attention Score of 125. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 February 2022.
All research outputs
#271,392
of 22,479,668 outputs
Outputs from Genome Biology (Online Edition)
#128
of 4,087 outputs
Outputs of similar age
#6,122
of 287,244 outputs
Outputs of similar age from Genome Biology (Online Edition)
#1
of 1 outputs
Altmetric has tracked 22,479,668 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 98th percentile: it's in the top 5% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 4,087 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 27.6. This one has done particularly well, scoring higher than 96% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 287,244 tracked outputs that were published within six weeks on either side of this one in any source. This one has done particularly well, scoring higher than 97% of its contemporaries.
We're also able to compare this research output to 1 others from the same source and published within six weeks on either side of this one. This one has scored higher than all of them