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Analysis of institutional authors

Pérez-Arnaiz P.AuthorAlmendros M.AuthorBerenguer J.AuthorHidalgo A.Author

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January 1, 2025
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Thermostable in vitro transcription-translation compatible with microfluidic droplets

Publicated to:Microbial Cell Factories. 23 169- - 2024-12-01 23(), DOI: 10.1186/s12934-024-02440-y

Authors: Ribeiro ALJL; Pérez-Arnaiz P; Sánchez-Costa M; Pérez L; Almendros M; van Vliet L; Gielen F; Lim J; Charnock S; Hollfelder F; González-Pastor JE; Berenguer J; Hidalgo A

Affiliations

CSIC-INTA - Centro de Astrobiologia (CAB) - Author
CSIC-UAM - Centro de Biología Molecular Severo Ochoa (CBM); Universidad Autónoma de Madrid - Author
CSIC-UAM - Centro de Biología Molecular Severo Ochoa (CBM); Universidad Autónoma de Madrid; Universidad Autónoma de Madrid - Author
DROP-TECH LTD; University of Exeter; University of Exeter - Author
Prozomix Limited - Author
University of Cambridge - Author
University of Cambridge; DROP-TECH LTD - Author
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Abstract

Background: In vitro expression involves the utilization of the cellular transcription and translation machinery in an acellular context to produce one or more proteins of interest and has found widespread application in synthetic biology and in pharmaceutical biomanufacturing. Most in vitro expression systems available are active at moderate temperatures, but to screen large libraries of natural or artificial genetic diversity for highly thermostable enzymes or enzyme variants, it is instrumental to enable protein synthesis at high temperatures. Objectives: Develop an in vitro expression system operating at high temperatures compatible with enzymatic assays and with technologies that enable ultrahigh-throughput protein expression in reduced volumes, such as microfluidic water-in-oil (w/o) droplets. Results: We produced cell-free extracts from Thermus thermophilus for in vitro translation including thermostable enzymatic cascades for energy regeneration and a moderately thermostable RNA polymerase for transcription, which ultimately limited the temperature of protein synthesis. The yield was comparable or superior to other thermostable in vitro expression systems, while the preparation procedure is much simpler and can be suited to different Thermus thermophilus strains. Furthermore, these extracts have enabled in vitro expression in microfluidic droplets at high temperatures for the first time. Conclusions: Cell-free extracts from Thermus thermophilus represent a simpler alternative to heavily optimized or pure component thermostable in vitro expression systems. Moreover, due to their compatibility with droplet microfluidics and enzyme assays at high temperatures, the reported system represents a convenient gateway for enzyme screening at higher temperatures with ultrahigh-throughput.

Keywords

Cell-free protein expressionDroplet microfluidicsIn vitro transcription and translationThermozymesThermus thermophiles

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Microbial Cell Factories due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2024 there are still no calculated indicators, but in 2023, it was in position 41/174, thus managing to position itself as a Q1 (Primer Cuartil), in the category Biotechnology & Applied Microbiology.

Independientemente del impacto esperado determinado por el canal de difusión, es importante destacar el impacto real observado de la propia aportación.

Según las diferentes agencias de indexación, el número de citas acumuladas por esta publicación hasta la fecha 2025-07-06:

  • Scopus: 1

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-07-06:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 9.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 9 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 2.75.
  • The number of mentions on the social network X (formerly Twitter): 6 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (Ribeiro A.L.J.L.) and Last Author (HIDALGO HUERTAS, AURELIO).