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Analysis of institutional authors

López-Márquez AAuthorMartínez-Pizarro AAuthorPerez BAuthorRichard EAuthorDesviat LrCorresponding Author

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March 25, 2022
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Modeling Splicing Variants Amenable to Antisense Therapy by Use of CRISPR-Cas9-Based Gene Editing in HepG2 Cells

Publicated to:Methods in Molecular Biology. 2434 167-184 - 2022-01-01 2434(), DOI: 10.1007/978-1-0716-2010-6_10

Authors: López-Márquez A; Martínez-Pizarro A; Pérez B; Richard E; Desviat LR

Affiliations

Centro de Biología Molecular Severo Ochoa CSIC-UAM, CEDEM, CIBERER (ISCIII), IdiPaz (ISCIII), Universidad Autónoma de Madrid, Madrid, Spain. - Author
Centro de Biología Molecular Severo Ochoa CSIC-UAM, CEDEM, CIBERER (ISCIII), IdiPaz (ISCIII), Universidad Autónoma de Madrid, Madrid, Spain. lruiz@cbm.csic.es. - Author

Abstract

The field of splice modulating RNA therapy has gained new momentum with FDA approved antisense-based drugs for several rare diseases. In vitro splicing assays with minigenes or patient-derived cells are commonly employed for initial preclinical testing of antisense oligonucleotides aiming to modulate splicing. However, minigenes do not include the full genomic context of the exons under study and patients' samples are not always available, especially if the gene is expressed solely in certain tissues (e.g. liver or brain). This is the case for specific inherited metabolic diseases such as phenylketonuria (PKU) caused by mutations in the liver-expressed PAH gene.Herein we describe the generation of mutation-specific hepatic cellular models of PKU using CRISPR/Cas9 system, which is a versatile and easy-to-use gene editing tool. We describe in detail the selection of the appropriate cell line, guidelines for design of RNA guides and donor templates, transfection procedures and growth and selection of single-cell colonies with the desired variant , which should result in the accurate recapitulation of the splicing defect.

Keywords

cellular modelscrispr/cas9gene editinghepg2inherited metabolic diseasesphenylketonuriaCellular modelsCrispr/cas9Gene editingHepg2Inherited metabolic diseasesPhenylketonuriaSplicing

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Methods in Molecular Biology, Q3 Agency Scopus (SJR), its regional focus and specialization in Genetics, give it significant recognition in a specific niche of scientific knowledge at an international level.

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 2025-07-18:

  • Google Scholar: 1
  • Scopus: 1
  • Europe PMC: 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 2025-07-18:

  • 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: 7.
  • 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: 7 (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: 4.65.
  • The number of mentions on the social network X (formerly Twitter): 7 (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.
  • Assignment of a Handle/URN as an identifier within the deposit in the Institutional Repository: https://repositorio.uam.es/handle/10486/709784

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 (LOPEZ MARQUEZ, ARISTIDES) and Last Author (RUIZ DESVIAT, LOURDES).

the author responsible for correspondence tasks has been RUIZ DESVIAT, LOURDES.