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| Open AccessPrecisely controlling endogenous protein dosage in hPSCs and derivatives to model FOXG1 syndrome
Altered dosage of developmental regulators such as transcription factors can result in disorders, such as FOXG1 syndrome. Here, the authors demonstrate the utility of SMASh technology for modulating protein dosage by modeling FOXG1 syndrome using human pluripotent stem cell-derived neurons and neural organoids.
- Wenliang Zhu
- , Boya Zhang
- & Baoyang Hu
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Article
| Open AccessA tetracycline-dependent ribozyme switch allows conditional induction of gene expression in Caenorhabditis elegans
Tools for conditional induction of gene expression in C. elegans are limited compared to other organisms. Here the authors present a tetracycline-dependent ribozyme that allows conditional control of a gene of interest.
- Lena A. Wurmthaler
- , Monika Sack
- & Martin Gamerdinger
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Article
| Open AccessEngineered bidirectional promoters enable rapid multi-gene co-expression optimization
Classic monodirectional promoters are of limited use for multiple gene co-expression. Here the authors generate a library of 168 bidirectional promoters for the yeast K. phaffii (syn. P. pastoris) with diverse expression profiles to optimize metabolic pathway design.
- Thomas Vogl
- , Thomas Kickenweiz
- & Anton Glieder
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Article
| Open AccessCarboxysome encapsulation of the CO2-fixing enzyme Rubisco in tobacco chloroplasts
Previous efforts to assemble Rubisco within a cyanobacterial carboxysome-derived protein shell in plant chloroplasts to concentrate CO2 have been unsuccessful. Here, Long et al. produce carboxysomes in tobacco chloroplasts that encapsulate the introduced Rubisco and enable autotrophic growth at elevated CO2.
- Benedict M. Long
- , Wei Yih Hee
- & G. Dean Price
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Article
| Open AccessSingle-pot glycoprotein biosynthesis using a cell-free transcription-translation system enriched with glycosylation machinery
The ability to produce homogeneous glycoproteins is expected to advance fundamental understanding in glycoscience, but current in vivo-based production systems have several limitations. Here, the authors develop an E. coli extract-based one-pot system for customized production of N-linked glycoproteins.
- Thapakorn Jaroentomeechai
- , Jessica C. Stark
- & Matthew P. DeLisa
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Article
| Open AccessVersatile and on-demand biologics co-production in yeast
The ability to combine the production of multiple biologics into a single ‘on demand’ system could help in situations where resources are limited. Here the authors demonstrate a proof-of-concept approach for the co-production of three biologics, allowing singular, mixed and combination drug products.
- Jicong Cao
- , Pablo Perez-Pinera
- & Timothy K. Lu
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Article
| Open AccessTuning the dynamic range of bacterial promoters regulated by ligand-inducible transcription factors
For synthetic gene circuits to behave as designed, ligand-inducible promoters should display predictable ON/OFF characteristics. Here the authors design multi-input hybrid promoters to build transcriptional logic gates.
- Ye Chen
- , Joanne M. L. Ho
- & Matthew R. Bennett
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| Open AccessA multiplexable TALE-based binary expression system for in vivo cellular interaction studies
Binary expression systems enable researchers to deliver loss-of-function or gain-of-function transgenes with spatial-temporal resolution in vivo. Here, the authors present a programmable TALE-based system for multiplexed orthogonal activation of transgenes in Drosophila.
- Markus Toegel
- , Ghows Azzam
- & Tudor A. Fulga
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| Open AccessEfficient protein production by yeast requires global tuning of metabolism
The contribution of metabolic pathways to protein secretion is largely unknown. Here, the authors find conserved metabolic patterns in yeast by examining genome-wide transcriptional responses in high protein secretion mutants and reveal critical factors that can be tuned for efficient protein secretion.
- Mingtao Huang
- , Jichen Bao
- & Jens Nielsen
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Article
| Open AccessPlant-made polio type 3 stabilized VLPs—a candidate synthetic polio vaccine
Despite the success of current vaccination against poliomyelitis, safe, cheap and effective vaccines remain sought for continuing eradication effort. Here the authors use plants to express stabilized virus-like particles of type 3 poliovirus that can induce a protective immune response in mice transgenic for the human poliovirus receptor.
- Johanna Marsian
- , Helen Fox
- & George P. Lomonossoff
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Article
| Open AccessEnhancing titres of therapeutic viral vectors using the transgene repression in vector production (TRiP) system
The maximum titre of therapeutic viral vectors can be adversely affected by the encoded transgene. Here the authors repress transgene expression in producing cells by employing the tryptophan RNA-binding attenuation protein and show that it improves titre of RNA- and DNA-based viral vectors expressing toxic transgenes.
- H. E. Maunder
- , J. Wright
- & D. C. Farley
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Article
| Open AccessAutomated optogenetic feedback control for precise and robust regulation of gene expression and cell growth
Optogenetics has emerged as a promising means to achieve gene expression control in bioprocess engineering, but current systems cannot respond to fluctuations in growth conditions. Here the authors overcome this limitation and develop an automated optogenetic feedback control system for precise and robust control of protein production in E. coli.
- Andreas Milias-Argeitis
- , Marc Rullan
- & Mustafa Khammash
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Article
| Open AccessImmunogenomic engineering of a plug-and-(dis)play hybridoma platform
Hybridomas are widely used for antibody screening and production due to their genetic stability and rapid proliferation. Here the authors demonstrate the rapid reprogramming of antibody specificity in hybridomas using CRISPR-Cas9.
- Mark Pogson
- , Cristina Parola
- & Sai T. Reddy
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Gene silencing by CRISPR interference in mycobacteria
Recombination-based tools for generating targeted mutations in Mycobacterium tuberculosislack efficiency. Here the authors present a CRISPR interference approach that is able to efficiently repress the expression of target genes in mycobacteria, in a rapid and cost-effective manner.
- Eira Choudhary
- , Preeti Thakur
- & Nisheeth Agarwal
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A plasmid-based multigene expression system for mammalian cells
Fluorescent proteins are widely used in molecular biology to visualize protein expression and localization. Here, Krizet al. describe an efficient and flexible modular plasmid-based eukaryotic expression cloning strategy for the homogeneous expression of several fluorescent proteins in one cell.
- Andrijana Kriz
- , Katharina Schmid
- & Philipp Berger