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A Medium-Throughput Single Cell CRISPR-Cas9 Assay to Assess Gene Essentiality

Overview of attention for article published in Biological Procedures Online, November 2015
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Title
A Medium-Throughput Single Cell CRISPR-Cas9 Assay to Assess Gene Essentiality
Published in
Biological Procedures Online, November 2015
DOI 10.1186/s12575-015-0028-4
Pubmed ID
Authors

A. R. Grassian, T. M. E. Scales, S. K. Knutson, K. W. Kuntz, N. J. McCarthy, C. E. Lowe, J. D. Moore, R. A. Copeland, H. Keilhack, J. J. Smith, J. A. Wickenden, S. Ribich

Abstract

Target selection for oncology is a crucial step in the successful development of therapeutics. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 editing of specific loci offers an alternative method to RNA interference and small molecule inhibitors for determining whether a cell line is dependent on a specific gene product for proliferation or survival. In our initial studies using CRISPR-Cas9 to verify the dependence on EZH2 activity for proliferation of a SMARCB1/SNF5/INI1 mutant malignant rhabdoid tumor (MRT) cell line, we noted that the initial reduction in proliferation was lost over time. We hypothesized that in the few cells that retain proliferative capacity, at least one allele of EZH2 remains functional. To verify this, we developed an assay to analyze 10s-100s of clonal cell populations for target gene disruption using restriction digest and fluorescent fragment length analyses. Our results clearly show that in cell lines in which EZH2 is essential for proliferation, at least one potentially functional allele of EZH2 is retained in the clones that survive. This assay clearly indicates whether or not a specific gene is essential for survival and/or proliferation in a given cell line. Such data can aid the development of more robust therapeutics by increasing confidence in target selection.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 30 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 30 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 5 17%
Student > Ph. D. Student 5 17%
Student > Bachelor 4 13%
Other 3 10%
Student > Postgraduate 3 10%
Other 5 17%
Unknown 5 17%
Readers by discipline Count As %
Agricultural and Biological Sciences 10 33%
Biochemistry, Genetics and Molecular Biology 7 23%
Medicine and Dentistry 5 17%
Pharmacology, Toxicology and Pharmaceutical Science 1 3%
Immunology and Microbiology 1 3%
Other 1 3%
Unknown 5 17%
Attention Score in Context

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 24 November 2015.
All research outputs
#19,944,091
of 25,373,627 outputs
Outputs from Biological Procedures Online
#147
of 192 outputs
Outputs of similar age
#199,540
of 292,406 outputs
Outputs of similar age from Biological Procedures Online
#3
of 4 outputs
Altmetric has tracked 25,373,627 research outputs across all sources so far. This one is in the 18th percentile – i.e., 18% of other outputs scored the same or lower than it.
So far Altmetric has tracked 192 research outputs from this source. They receive a mean Attention Score of 4.6. This one is in the 19th percentile – i.e., 19% 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 292,406 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 26th percentile – i.e., 26% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 4 others from the same source and published within six weeks on either side of this one.