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Recombinant Protein Expression in Mammalian Cells

Overview of attention for book
Cover of 'Recombinant Protein Expression in Mammalian Cells'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 Transient Gene Expression in Suspension HEK293-EBNA1 Cells
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    Chapter 2 Transient Expression of Recombinant Membrane-eGFP Fusion Proteins in HEK293 Cells
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    Chapter 3 PEI-Mediated Transient Gene Expression in CHO Cells
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    Chapter 4 Stable Expression by Lentiviral Transduction of Cells
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    Chapter 5 Inducible Protein Production in 293 Cells Using the piggyBac Transposon System
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    Chapter 6 Recombinant CHO Cell Pool Generation Using piggyBac Transposon System
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    Chapter 7 Genome Engineering of Hybridomas to Generate Stable Cell Lines for Antibody Expression
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    Chapter 8 Protein Expression via Transient Transfection of Mammalian Cells in a WAVE Bioreactor
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    Chapter 9 CHO and HEK293 Cultivation and Transfection in Single-Use Orbitally Shaken Bioreactors
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    Chapter 10 Bench-Scale Stirred-Tank Bioreactor for Recombinant Protein Production in Chinese Hamster Ovary (CHO) Cells in Suspension
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    Chapter 11 Continuous and Integrated Expression and Purification of Recombinant Antibodies
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    Chapter 12 High Throughput Transfection of HEK293 Cells for Transient Protein Production
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    Chapter 13 Microfluidic Transfection for High-Throughput Mammalian Protein Expression
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    Chapter 14 Genome-Wide High-Throughput RNAi Screening for Identification of Genes Involved in Protein Production
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    Chapter 15 Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells
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    Chapter 16 Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells
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    Chapter 17 Scalable Production and Purification of Adeno-Associated Viral Vectors (AAV)
  19. Altmetric Badge
    Chapter 18 Considerations in the Use of Codon Optimization for Recombinant Protein Expression
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    Chapter 19 Versatile Cell-Free Protein Synthesis Systems Based on Chinese Hamster Ovary Cells
Attention for Chapter 15: Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells
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About this Attention Score

  • Above-average Attention Score compared to outputs of the same age (53rd percentile)
  • High Attention Score compared to outputs of the same age and source (80th percentile)

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Chapter title
Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells
Chapter number 15
Book title
Recombinant Protein Expression in Mammalian Cells
Published in
Methods in molecular biology, September 2018
DOI 10.1007/978-1-4939-8730-6_15
Pubmed ID
Book ISBNs
978-1-4939-8729-0, 978-1-4939-8730-6
Authors

Kevin Kellner, Ankur Solanki, Thomas Amann, Nga Lao, Niall Barron, Kellner, Kevin, Solanki, Ankur, Amann, Thomas, Lao, Nga, Barron, Niall

Abstract

MicroRNAs with their unique ability to target hundreds of genes have been highlighted as powerful tools to improve bioprocess behavior of cells. The common approaches to stably deplete miRNAs are the use of sponge decoy transcripts or shRNA inhibitors, which requires the introduction and expression of extra genetic material. As an alternative, we implemented the CRISPR/Cas9 system in our laboratory to generate Chinese hamster ovary (CHO) cells which lack the expression of a specific miRNA for the purpose of functional studies. To implement the system, miR-27a/b was chosen as it has been shown to be upregulated during hypothermic conditions and therefore may be involved in controlling CHO cell growth and recombinant protein productivity. In this chapter, we present a protocol for targeting miRNAs in CHO cells using CRISPR/Cas9 and the analysis of the resulting phenotype, using miR-27 as an example. We showed that it is possible to target miRNAs in CHO cells and achieved ≥80% targeting efficiency. Indel analysis and TOPO-TA cloning combined with Sanger sequencing showed a range of different indels. Furthermore, it was possible to identify clones with no detectable expression of mature miR-27b. Depletion of miR-27b led to improved viability in late stages of batch and fed-batch cultures making it a potentially interesting target to improve bioprocess performance of CHO cells.

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X Demographics

The data shown below were collected from the profiles of 5 X users who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 24 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 24 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 6 25%
Researcher 4 17%
Student > Master 2 8%
Student > Doctoral Student 1 4%
Professor 1 4%
Other 3 13%
Unknown 7 29%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 12 50%
Agricultural and Biological Sciences 2 8%
Medicine and Dentistry 1 4%
Unknown 9 38%
Attention Score in Context

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 21 August 2023.
All research outputs
#8,095,869
of 24,294,745 outputs
Outputs from Methods in molecular biology
#2,509
of 13,669 outputs
Outputs of similar age
#136,271
of 344,478 outputs
Outputs of similar age from Methods in molecular biology
#38
of 236 outputs
Altmetric has tracked 24,294,745 research outputs across all sources so far. This one is in the 44th percentile – i.e., 44% of other outputs scored the same or lower than it.
So far Altmetric has tracked 13,669 research outputs from this source. They receive a mean Attention Score of 3.5. This one has done well, scoring higher than 75% 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 344,478 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 53% of its contemporaries.
We're also able to compare this research output to 236 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 80% of its contemporaries.