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Grant support

We thank all members of our laboratories for their contributions and helpful discussions. We thank the CRG Genomics Unit and the Biomolecular Screening & Protein Technologies Unit. We acknowledge help in the quantifications of the Western blots by Dina Cramer and assistance in RNA sequencing analysis by Javier Delgado. Armelle Yart provided the HEK293 cells stably expressing GH receptor. This work was funded by the European Commission (EC) Framework programme (FP) 7 projects PRIMES (contract nr. 278568), ComplexINC (contract nr. 279039) and SynSignal (contract nr. 613879). LS is supported by the Spanish Ministerio de Economia y Competitividad, Plan Nacional BIO2012-39754 and the European Fund for Economic and Regional Development. We are particularly grateful for the support of the Spanish Ministry of Economy and Competitiveness, 'Centro de Excelencia Severo Ochoa 2013-2017' (SEV-2012-0208).

Analysis of institutional authors

Beltran-Sastre VAuthorBenisty HAuthorBurnier JAuthorSerrano LCorresponding AuthorKiel CAuthor

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Article

Tuneable endogenous mammalian target complementation via multiplexed plasmid-based recombineering

Publicated to:Scientific Reports. 5 17432- - 2015-11-27 5(), DOI: 10.1038/srep17432

Authors: Beltran-Sastre, V; Benisty, H; Burnier, J; Berger, I; Serrano, L; Kiel, C

Affiliations

CRG, EMBL CRG Syst Biol Res Unit, Barcelona 08003, Spain - Author
European Mol Biol Lab, F-38042 Grenoble, France - Author
ICREA, Barcelona 08010, Spain - Author
Univ Bristol, Sch Biochem, Clifton BS8 1TD, England - Author
UPF, Barcelona 08003, Spain - Author
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Abstract

Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce 'TEMTAC', a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.

Keywords

AnimalsBraf protein, humanCellsDnaExpressionGene silencingGene transfer techniquesGenes, reporterGenetic engineeringGenetic vectorsHek293 cellsHela cellsHras protein, humanHumansLuciferasesMelanomaMutationPlasmidsProtein tyrosine phosphatase, non-receptor type 11Proto-oncogene proteins b-rafProto-oncogene proteins p21(ras)RnaRna, small interferingSystemTissuesTranscription

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Scientific Reports 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, 2015, it was in position 7/62, thus managing to position itself as a Q1 (Primer Cuartil), in the category Multidisciplinary Sciences.

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 2025-06-20:

  • WoS: 3
  • Scopus: 3
  • Europe PMC: 2
  • OpenCitations: 4

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 2025-06-20:

  • 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: 15.
  • 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: 15 (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: 1.
  • The number of mentions on the social network X (formerly Twitter): 1 (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

This work has been carried out with international collaboration, specifically with researchers from: France; United Kingdom.

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 (BELTRÁN SASTRE, VIOLETA) and Last Author (Kiel, CHRISTINA).

the author responsible for correspondence tasks has been SERRANO PUBUL, LUIS.