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A lentivirus-free inducible CRISPR-Cas9 system for efficient targeting of human genes

Overview of attention for article published in Analytical Biochemistry, August 2017
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About this Attention Score

  • Good Attention Score compared to outputs of the same age (68th percentile)
  • High Attention Score compared to outputs of the same age and source (93rd percentile)

Mentioned by

twitter
4 tweeters
patent
1 patent

Citations

dimensions_citation
5 Dimensions

Readers on

mendeley
53 Mendeley
Title
A lentivirus-free inducible CRISPR-Cas9 system for efficient targeting of human genes
Published in
Analytical Biochemistry, August 2017
DOI 10.1016/j.ab.2017.05.001
Pubmed ID
Authors

Kamlesh Bisht, Sherilyn Grill, Jacqueline Graniel, Jayakrishnan Nandakumar

Abstract

CRISPR-Cas9 is a cutting-edge tool for modifying genomes. The efficacy with which Cas9 recognizes its target has revolutionized the engineering of knockouts. However this efficacy complicates the knocking out of important genes in cultured cells. Unedited cells holding a survival advantage within an edited population can confound the knockout phenotype. Here we develop a HeLa-based system that overcomes this limitation, incorporating several attractive features. First, we use Flp-recombinase to generate clones stably integrated for Cas9 and guide RNAs, eliminating the possibility of unedited cells. Second, Cas9 can be induced uniformly in the clonal cultures using doxycycline to measure the knockout phenotype. Third, two genes can be simultaneously knocked out using this approach. Finally, by not involving lentiviruses, our method is appealing to a broad research audience. Using this methodology we generated an inducible AGO2-knockout cell line showing normal RNA interference in the absence of doxycycline. Upon induction of Cas9, the AGO2 locus was cleaved, the AGO2 protein was depleted, and RNA interference was compromised. In addition to generating inducible knockouts, our technology can be adapted to improve other applications of Cas9, including transcriptional/epigenetic modulation and visualization of cellular DNA loci.

Twitter Demographics

The data shown below were collected from the profiles of 4 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 53 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Italy 1 2%
Unknown 52 98%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 15 28%
Researcher 12 23%
Student > Bachelor 10 19%
Student > Postgraduate 3 6%
Student > Doctoral Student 3 6%
Other 6 11%
Unknown 4 8%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 27 51%
Agricultural and Biological Sciences 15 28%
Medicine and Dentistry 4 8%
Pharmacology, Toxicology and Pharmaceutical Science 1 2%
Arts and Humanities 1 2%
Other 0 0%
Unknown 5 9%

Attention Score in Context

This research output has an Altmetric Attention Score of 5. 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 20 December 2018.
All research outputs
#3,923,410
of 14,834,764 outputs
Outputs from Analytical Biochemistry
#2,137
of 7,236 outputs
Outputs of similar age
#82,341
of 265,367 outputs
Outputs of similar age from Analytical Biochemistry
#2
of 32 outputs
Altmetric has tracked 14,834,764 research outputs across all sources so far. This one has received more attention than most of these and is in the 73rd percentile.
So far Altmetric has tracked 7,236 research outputs from this source. They receive a mean Attention Score of 4.0. This one has gotten more attention than average, scoring higher than 70% 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 265,367 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 68% of its contemporaries.
We're also able to compare this research output to 32 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 93% of its contemporaries.