Expression systems

  • Article
    | Open Access

    Chimeric antigen receptors (CAR) use antibody variable regions to activate anti-tumor immunity. Here the authors show that a mouse IgH/IgL variable region used in a clinical CAR induces host immune responses to possibly reduce therapy efficacy, but an IgH-only CAR T design achieves similar CAR T activity but is potentially less immunogenic.

    • Norris Lam
    • , Nathan D. Trinklein
    • , Benjamin Buelow
    • , George H. Patterson
    • , Namrata Ojha
    •  & James N. Kochenderfer
  • Article
    | Open Access

    Several factors contribute to the efficiency of protein expression. Here the authors show that the identity of amino acids encoded by codons at position 3–5 significantly impact translation efficiency and protein expression levels.

    • Manasvi Verma
    • , Junhong Choi
    • , Kyle A. Cottrell
    • , Zeno Lavagnino
    • , Erica N. Thomas
    • , Slavica Pavlovic-Djuranovic
    • , Pawel Szczesny
    • , David W. Piston
    • , Hani S. Zaher
    • , Joseph D. Puglisi
    •  & Sergej Djuranovic
  • Article
    | Open Access

    Sustainable sources are needed to meet the demand for long-chain polyunsaturated fatty acids. Here the authors construct an artificial biosynthetic gene cluster in Y. lipolytica capable of producing a high concentration of PUFAs.

    • Katja Gemperlein
    • , Demian Dietrich
    • , Michael Kohlstedt
    • , Gregor Zipf
    • , Hubert S. Bernauer
    • , Christoph Wittmann
    • , Silke C. Wenzel
    •  & Rolf Müller
  • Article
    | Open Access

    Artificial cells need to be supplied with ATP as they lack internal systems of energy generation. Here the authors reconstruct ATP synthase and bacteriorhodopsins for light-driven ATP generation, powering transcription and translation.

    • Samuel Berhanu
    • , Takuya Ueda
    •  & Yutetsu Kuruma
  • Article
    | Open Access

    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
    • , Mengqi Li
    • , Fan Mo
    • , Tingwei Mi
    • , Yihui Wu
    • , Zhaoqian Teng
    • , Qi Zhou
    • , Wei Li
    •  & Baoyang Hu
  • Article
    | Open Access

    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
    • , Julia Pitzer
    • , Lukas Sturmberger
    • , Astrid Weninger
    • , Bradley W. Biggs
    • , Eva-Maria Köhler
    • , Armin Baumschlager
    • , Jasmin Elgin Fischer
    • , Patrick Hyden
    • , Marlies Wagner
    • , Martina Baumann
    • , Nicole Borth
    • , Martina Geier
    • , Parayil Kumaran Ajikumar
    •  & Anton Glieder
  • Article
    | Open Access

    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
    • , Robert E. Sharwood
    • , Benjamin D. Rae
    • , Sarah Kaines
    • , Yi-Leen Lim
    • , Nghiem D. Nguyen
    • , Baxter Massey
    • , Soumi Bala
    • , Susanne von Caemmerer
    • , Murray R. Badger
    •  & G. Dean Price
  • Article
    | Open Access

    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
    • , Aravind Natarajan
    • , Cameron J. Glasscock
    • , Laura E. Yates
    • , Karen J. Hsu
    • , Milan Mrksich
    • , Michael C. Jewett
    •  & Matthew P. DeLisa
  • Article
    | Open Access

    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
    • , Ky Lowenhaupt
    • , Ming-Ru Wu
    • , Oliver Purcell
    • , Cesar de la Fuente-Nunez
    •  & Timothy K. Lu
  • Article
    | Open Access

    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
    • , Eunice Y. Lee
    • , David J. H. F. Knapp
    • , Ying Tan
    • , Ming Fa
    •  & Tudor A. Fulga
  • Article
    | Open Access

    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
    • , Björn M. Hallström
    • , Dina Petranovic
    •  & Jens Nielsen
  • Article
    | Open Access

    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
    • , Mohammad W. Bahar
    • , Abhay Kotecha
    • , Elizabeth E. Fry
    • , David I. Stuart
    • , Andrew J. Macadam
    • , David J. Rowlands
    •  & George P. Lomonossoff
  • Article
    | Open Access

    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
    • , B. R. Kolli
    • , C. R. Vieira
    • , T. T. Mkandawire
    • , S. Tatoris
    • , V. Kennedy
    • , S. Iqball
    • , G. Devarajan
    • , S. Ellis
    • , Y. Lad
    • , N. G. Clarkson
    • , K. A. Mitrophanous
    •  & D. C. Farley
  • Article
    | Open Access

    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
    • , Stephanie K. Aoki
    • , Peter Buchmann
    •  & Mustafa Khammash
  • Article
    | Open Access

    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
    • , William J. Kelton
    • , Paul Heuberger
    •  & Sai T. Reddy
  • Article |

    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
    • , Madhu Pareek
    •  & Nisheeth Agarwal
  • Article |

    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
    • , Nadia Baumgartner
    • , Urs Ziegler
    • , Imre Berger
    • , Kurt Ballmer-Hofer
    •  & Philipp Berger