News Improve Your Protein and Virus Production

Improve Your Protein and Virus Production

November 21, 2017

Quantitation of fat metabolism in individual adipocytes from multiple patients is no longer as tedious as finding a needle in a haystack thanks to high-content/highthroughput microscopy analysis of lipid droplets. Read the accompanying article by Bombrun and coworkers in this issue of Cytometry Part A.

ALSO IN THE NOVEMBER ISSUE

Generation of apoptosis-resistant HEK293 cells using CRISPR/Cas gene-editing tool for improved protein and virus production.

Apoptosis has important functions during pathophysiologic processes. However, from a biopharmaceutical point of view, active apoptosis of host cells is undesirable during viral packaging or protein expression, because it decreases the efficiency of viral or protein production. In this Biotechnology and Bioengineering article, Zhang and coworkers established an apoptosis-resistant HEK293 cells with CRISPR/Cas mediated quadruple gene knockout to improve protein and virus production. Caspase3 gene was firstly knocked out in HEK293 cells followed by the simultaneous knockout of Caspase6, Caspase7 and AIF1 genes. The produced cell line is proven to be resistant to apoptosis triggers from both intracellular and extracellular environments, and shows increased capacity of protein expression and viral packaging.

Zhang et al. Biotechnol Bioeng 2017; 114. DOI: 10.1002/bit.26382

A strong hybridfatty acid inducible transcriptional sensor built from Yarrowia lipolytica upstream activating and regulatory sequences

Microbial systems are regularly engineered to produce biochemicals, materials, and therapeutics. The engineering of Yarrowia lipolytica to accumulate lipids with high titers and productivities has been enabled with a handful of constitutive promoters for pathway  engineering. A strong and tightly regulated fatty acid inducible promoter for Yarrowia lipolytica that could enable more advanced metabolic control strategies is developed. In this Biotechnology Journal article, Hussain and coworkers engineer a fatty acid regulated hybrid  promoter for use in Y. lipolytica. A 200 bp upstream regulatory sequence in the peroxisomal acyl CoA oxidase 2 (POX2) promoter is identified. Further analysis of the promoter sequence reveal a regulatory sequence, that when used in tandem repeats, lead to a 48-fold induction of gene expression relative to glucose and fourfold higher than the native POX2 promoter. To date, this is the strongest inducible promoter reported in Y. lipolytica. Taken together, the results show that it is possible to engineer strong promoters that retain strong inducibility. These types of promoters will be useful in controlling metabolism and as fatty acid sensors.

Hussain et al. Biotechnol J 2017; 12 . DOI: 10.1002/ biot.201700248

 


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