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Engineering of Primary Human B cells with CRISPR/Cas9 Targeted Nuclease

Overview of attention for article published in Scientific Reports, August 2018
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About this Attention Score

  • Above-average Attention Score compared to outputs of the same age (60th percentile)
  • High Attention Score compared to outputs of the same age and source (90th percentile)

Mentioned by

twitter
8 tweeters

Citations

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7 Dimensions

Readers on

mendeley
63 Mendeley
Title
Engineering of Primary Human B cells with CRISPR/Cas9 Targeted Nuclease
Published in
Scientific Reports, August 2018
DOI 10.1038/s41598-018-30358-0
Pubmed ID
Authors

Matthew J. Johnson, Kanut Laoharawee, Walker S. Lahr, Beau R. Webber, Branden S. Moriarity

Abstract

B cells offer unique opportunities for gene therapy because of their ability to secrete large amounts of protein in the form of antibody and persist for the life of the organism as plasma cells. Here, we report optimized CRISPR/Cas9 based genome engineering of primary human B cells. Our procedure involves enrichment of CD19+ B cells from PBMCs followed by activation, expansion, and electroporation of CRISPR/Cas9 reagents. We are able expand total B cells in culture 10-fold and outgrow the IgD+ IgM+ CD27- naïve subset from 35% to over 80% of the culture. B cells are receptive to nucleic acid delivery via electroporation 3 days after stimulation, peaking at Day 7 post stimulation. We tested chemically modified sgRNAs and Alt-R gRNAs targeting CD19 with Cas9 mRNA or Cas9 protein. Using this system, we achieved genetic and protein knockout of CD19 at rates over 70%. Finally, we tested sgRNAs targeting the AAVS1 safe harbor site using Cas9 protein in combination with AAV6 to deliver donor template encoding a splice acceptor-EGFP cassette, which yielded site-specific integration frequencies up to 25%. The development of methods for genetically engineered B cells opens the door to a myriad of applications in basic research, antibody production, and cellular therapeutics.

Twitter Demographics

The data shown below were collected from the profiles of 8 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 63 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 63 100%

Demographic breakdown

Readers by professional status Count As %
Unspecified 15 24%
Researcher 15 24%
Student > Ph. D. Student 8 13%
Student > Master 6 10%
Student > Bachelor 6 10%
Other 13 21%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 18 29%
Unspecified 17 27%
Agricultural and Biological Sciences 13 21%
Medicine and Dentistry 5 8%
Immunology and Microbiology 4 6%
Other 6 10%

Attention Score in Context

This research output has an Altmetric Attention Score of 3. 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 29 August 2018.
All research outputs
#3,652,671
of 12,559,973 outputs
Outputs from Scientific Reports
#21,207
of 57,931 outputs
Outputs of similar age
#104,406
of 273,080 outputs
Outputs of similar age from Scientific Reports
#1
of 11 outputs
Altmetric has tracked 12,559,973 research outputs across all sources so far. This one is in the 49th percentile – i.e., 49% of other outputs scored the same or lower than it.
So far Altmetric has tracked 57,931 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 15.3. This one has gotten more attention than average, scoring higher than 62% 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 273,080 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 60% of its contemporaries.
We're also able to compare this research output to 11 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 90% of its contemporaries.