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Highly efficient generation of knock-in transgenic medaka by CRISPR/Cas9-mediated genome engineering

Overview of attention for article published in Zoological Letters, February 2018
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Title
Highly efficient generation of knock-in transgenic medaka by CRISPR/Cas9-mediated genome engineering
Published in
Zoological Letters, February 2018
DOI 10.1186/s40851-017-0086-3
Pubmed ID
Authors

Ikuko Watakabe, Hisashi Hashimoto, Yukiko Kimura, Saori Yokoi, Kiyoshi Naruse, Shin-ichi Higashijima

Abstract

Medaka (Oryzias latipes) is a popular animal model used in vertebrate genetic analysis. Recently, an efficient (~ 30%) knock-in system via non-homologous end joining (NHEJ) was established in zebrafish using the CRISPR/Cas9 system. If the same technique were applicable in medaka, it would greatly expand the usefulness of this model organism. The question of the applicability of CRISPR/Cas9 in medaka, however, has yet to be addressed. We report the highly efficient generation of knock-in transgenic medaka via non-homologous end joining (NHEJ). Donor plasmid containing a heat-shock promoter and a reporter gene was co-injected with a short guide RNA (sgRNA) targeted for genome digestion, an sgRNA targeted for donor plasmid digestion, and Cas9 mRNA. Broad transgene expression in the expression domain of a target gene was observed in approximately 25% of injected embryos. By raising these animals, we established stable knock-in transgenic fish with several different constructs for five genetic loci, obtaining transgenic founders at efficiencies of > 50% for all five loci. Further, we show that the method is useful for obtaining mutant alleles. In the experiments where transgene integrations were targeted between the transcription start site and the initiation methionine, the resultant transgenic fish became mutant alleles. With its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in via NHEJ will become a standard method for the generation of transgenic and mutant medaka.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 118 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 118 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 24 20%
Student > Ph. D. Student 15 13%
Student > Bachelor 15 13%
Student > Master 10 8%
Student > Doctoral Student 7 6%
Other 12 10%
Unknown 35 30%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 30 25%
Agricultural and Biological Sciences 25 21%
Neuroscience 9 8%
Medicine and Dentistry 4 3%
Environmental Science 3 3%
Other 7 6%
Unknown 40 34%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. 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 16 February 2018.
All research outputs
#14,091,602
of 23,020,670 outputs
Outputs from Zoological Letters
#124
of 169 outputs
Outputs of similar age
#231,302
of 437,337 outputs
Outputs of similar age from Zoological Letters
#4
of 5 outputs
Altmetric has tracked 23,020,670 research outputs across all sources so far. This one is in the 37th percentile – i.e., 37% of other outputs scored the same or lower than it.
So far Altmetric has tracked 169 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 13.7. This one is in the 25th percentile – i.e., 25% of its peers scored the same or lower than it.
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 437,337 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 45th percentile – i.e., 45% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 5 others from the same source and published within six weeks on either side of this one.