Evidence map›Paper›PMID 32703203›Full record

ArticleMicrobial cell factories2020

A novel knock out strategy to enhance recombinant protein expression in Escherichia coli.

Ashish K Sharma, Esha Shukla, Deepak S Janoti, Krishna J Mukherjee, Joseph Shiloach

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Microbial cell factories, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.

0numbers the graph read from it
0cells of the map it votes in
12citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

12 citing papers in PubMed.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Ashish K SharmaSchool of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
Esha ShuklaSchool of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
Deepak S JanotiSchool of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
Krishna J MukherjeeSchool of Biotechnology, Jawaharlal Nehru University, New Delhi, India. kjmukherjee@jnu.ac.in.
Joseph ShiloachBiotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health, Bethesda, MD, 20892, USA. josephs@niddk.nih.gov.ORCID http://orcid.org/0000-0002-9678-6778

Funding

Understanding metabolism and stress conditions of recombinant microorganismsZIADK075081 · NIDDK · NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES · PI SHILOACH, JOSEPH · 2012 to 2022
$2.6M
NIDDK NIH HHS DK075081
6 · The paper itself

Abstract

backgroundThe expression of recombinant proteins triggers a stress response which downregulates key metabolic pathway genes leading to a decline in cellular health and feedback inhibition of both growth and protein expression. Instead of individually upregulating these downregulated genes or improving transcription rates by better vector design, an innovative strategy would be to block this stress response thereby ensuring a sustained level of protein expression.

resultsWe postulated that the genes which are commonly up-regulated post induction may play the role of signalling messengers in mounting the cellular stress response. We identified those genes which have no known downstream regulatees and created knock outs which were then tested for GFP expression. Many of these knock outs showed significantly higher expression levels which was also sustained for longer periods. The highest product yield (Y

conclusionCreating key knock outs to block the CSR and enhance expression is a radically different strategy that can be synergistically combined with traditional methods of improving protein yields thus helping in the design of superior host platforms for protein expression.

Indexed as

AsparaginaseEscherichia coliEscherichia coli ProteinsFermentationGene Expression Regulation, BacterialGene Knockout TechniquesGenes, BacterialGreen Fluorescent ProteinsMetabolic Networks and PathwaysRecombinant ProteinsSignal TransductionStress, PhysiologicalUp-RegulationAsparaginaseEscherichia coli ProteinsGreen Fluorescent ProteinsRecombinant ProteinsCellular stress responseEscherichia coliKnock outRecombinant protein expressionSignalling control

Identifiers

PMID32703203
PMCPMC7376861

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.