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enChIP systems using different CRISPR orthologues and epitope tags

Overview of attention for article published in BMC Research Notes, February 2018
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Title
enChIP systems using different CRISPR orthologues and epitope tags
Published in
BMC Research Notes, February 2018
DOI 10.1186/s13104-018-3262-4
Pubmed ID
Authors

Toshitsugu Fujita, Miyuki Yuno, Hodaka Fujii

Abstract

Previously, we developed the engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) technology, which isolates specific genomic regions while preserving their molecular interactions. In enChIP, the locus of interest is tagged with engineered DNA-binding molecules such as the clustered regularly interspaced short palindromic repeats (CRISPR) system, consisting of a catalytically inactive form of Cas9 (dCas9) and guide RNA, followed by affinity purification of the tagged locus to allow identification of associated molecules. In our previous studies, we used a 3xFLAG-tagged CRISPR system from Streptococcus pyogenes (S. pyogenes). In this study, to increase the flexibility of enChIP, we used the CRISPR system from Staphylococcus aureus (S. aureus) along with different epitope tags. We generated a plasmid expressing S. aureus dCas9 (Sa-dCas9) fused to a nuclear localization signal (NLS) and a 3xFLAG-tag (Sa-dCas9-3xFLAG). The yields of enChIP using Sa-dCas9-3xFLAG were comparable to those using S. pyogenes dCas9 fused with an NLS and a 3xFLAG-tag (3xFLAG-Sp-dCas9). We also generated another enChIP system using Sp-dCas9 fused with an NLS and a 2xAM-tag (Sp-dCas9-2xAM). We obtained high enChIP yields using this system as well. Our findings indicate that these tools will increase the flexibility of enChIP analysis.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 37 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 37 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 7 19%
Student > Master 6 16%
Researcher 5 14%
Student > Bachelor 5 14%
Professor 3 8%
Other 6 16%
Unknown 5 14%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 16 43%
Agricultural and Biological Sciences 10 27%
Pharmacology, Toxicology and Pharmaceutical Science 3 8%
Medicine and Dentistry 2 5%
Neuroscience 1 3%
Other 0 0%
Unknown 5 14%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. 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 04 March 2018.
All research outputs
#11,208,038
of 12,593,681 outputs
Outputs from BMC Research Notes
#2,356
of 2,835 outputs
Outputs of similar age
#236,454
of 272,707 outputs
Outputs of similar age from BMC Research Notes
#1
of 1 outputs
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