High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing

The CYP21A2 gene, mapped to the RCCX module in 6p21.3, is responsible for 21-hydroxylase deficiency (21-HD). In this work, we leveraged Oxford Nanopore Technology (ONT) Long Read sequencing (LRS) to analyze samples from an Argentinian cohort of 21-HD. A total of 34 samples were sequenced in 2 amp...

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Main Authors: Claps, Aldana, Fernández, Cecilia, Fernandez, Franco Daniel, Macchiaroli, Natalia, Ingravidi, Marina L., Delea, Marisol, Castro, Tania, Laiseca, Julieta, Kamenetzky, Laura, Taboas, Melisa, Dain, Liliana
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Springer Nature 2025
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/23066
https://www.nature.com/articles/s41598-025-03799-7
https://doi.org/10.1038/s41598-025-03799-7
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author Claps, Aldana
Fernández, Cecilia
Fernandez, Franco Daniel
Macchiaroli, Natalia
Ingravidi, Marina L.
Delea, Marisol
Fernández, Cecilia
Castro, Tania
Laiseca, Julieta
Kamenetzky, Laura
Taboas, Melisa
Dain, Liliana
author_browse Castro, Tania
Claps, Aldana
Dain, Liliana
Delea, Marisol
Fernandez, Franco Daniel
Fernández, Cecilia
Ingravidi, Marina L.
Kamenetzky, Laura
Laiseca, Julieta
Macchiaroli, Natalia
Taboas, Melisa
author_facet Claps, Aldana
Fernández, Cecilia
Fernandez, Franco Daniel
Macchiaroli, Natalia
Ingravidi, Marina L.
Delea, Marisol
Fernández, Cecilia
Castro, Tania
Laiseca, Julieta
Kamenetzky, Laura
Taboas, Melisa
Dain, Liliana
author_sort Claps, Aldana
collection INTA Digital
description The CYP21A2 gene, mapped to the RCCX module in 6p21.3, is responsible for 21-hydroxylase deficiency (21-HD). In this work, we leveraged Oxford Nanopore Technology (ONT) Long Read sequencing (LRS) to analyze samples from an Argentinian cohort of 21-HD. A total of 34 samples were sequenced in 2 amplicons of 8.5 Kb covering the centromeric and telomeric RCCX modules. The number of variants found varied between 3 and 106 and all expected pathogenic variants and new ones were obtained with the LR sequencing workflow developed. We defined with higher accuracy the breakpoints of the rearrangements allowing the reclassification of chimeras and/or converted genes in 18.75% of the samples, some of them with clinical implications. By addressing the study of the telomeric RCCX module/s in depth, we found 19 genetic variants (GVs) for CYP21A1P and 29 GVs for TNXA not previously described in Latin American populations. This study may represent the first application of ONT LRS for clinical evaluation of Latin American subjects, highlighting the importance of LRS as a high-resolution method of diagnosis. It would allow a better understanding of the diversity of the RCCX modules and improve our knowledge of the variation of genetic mechanisms behind the disease.
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spelling INTA230662025-07-17T14:49:48Z High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing Claps, Aldana Fernández, Cecilia Fernandez, Franco Daniel Macchiaroli, Natalia Ingravidi, Marina L. Delea, Marisol Fernández, Cecilia Castro, Tania Laiseca, Julieta Kamenetzky, Laura Taboas, Melisa Dain, Liliana Secuenciación por Nanoporos Genética Nanopore Sequencing Genetics RCCX Modules CYP21A2 Long Read Sequencing 21-hydroxylase Deficiency The CYP21A2 gene, mapped to the RCCX module in 6p21.3, is responsible for 21-hydroxylase deficiency (21-HD). In this work, we leveraged Oxford Nanopore Technology (ONT) Long Read sequencing (LRS) to analyze samples from an Argentinian cohort of 21-HD. A total of 34 samples were sequenced in 2 amplicons of 8.5 Kb covering the centromeric and telomeric RCCX modules. The number of variants found varied between 3 and 106 and all expected pathogenic variants and new ones were obtained with the LR sequencing workflow developed. We defined with higher accuracy the breakpoints of the rearrangements allowing the reclassification of chimeras and/or converted genes in 18.75% of the samples, some of them with clinical implications. By addressing the study of the telomeric RCCX module/s in depth, we found 19 genetic variants (GVs) for CYP21A1P and 29 GVs for TNXA not previously described in Latin American populations. This study may represent the first application of ONT LRS for clinical evaluation of Latin American subjects, highlighting the importance of LRS as a high-resolution method of diagnosis. It would allow a better understanding of the diversity of the RCCX modules and improve our knowledge of the variation of genetic mechanisms behind the disease. Instituto de Patología Vegetal Fil: Claps, Aldana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Centro Nacional de Genética Médica; Argentina Fil: Fernández, Cecilia. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3); Argentina Fil: Fernandez, Franco Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Fernandez, Franco Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Macchiaroli, Natalia. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3); Argentina Fil: Ingravidi, Marina L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3); Argentina Fil: Delea, Marisol. Hospital de Alta Complejidad El Calafate SAMIC. Unidad de Conocimiento Traslacional Hospitalaria Patagónica; Argentina Fil: Fernández, Cecilia. Laboratorio Novagen; Argentina Fil: Castro, Tania. Administración Nacional de Laboratorios e Institutos de Salud (ANLIS) “Dr. Carlos G Malbrán”. Centro Nacional de Genética Médica; Argentina Fil: Laiseca, Julieta. Administración Nacional de Laboratorios e Institutos de Salud (ANLIS) “Dr. Carlos G Malbrán”. Centro Nacional de Genética Médica; Argentina Fil: Kamenetzky, Laura. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3); Argentina Fil: Taboas, Melisa. Administración Nacional de Laboratorios e Institutos de Salud (ANLIS) “Dr. Carlos G Malbrán”. Centro Nacional de Genética Médica; Argentina Fil: Dain, Liliana. Administración Nacional de Laboratorios e Institutos de Salud (ANLIS) “Dr. Carlos G Malbrán. Centro Nacional de Genética Médica; Argentina Fil: Dain, Liliana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3); Argentina 2025-07-17T14:43:25Z 2025-07-17T14:43:25Z 2025-07-10 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/23066 https://www.nature.com/articles/s41598-025-03799-7 2045-2322 https://doi.org/10.1038/s41598-025-03799-7 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Springer Nature Scientific Reports 15 : 24983 (July 2025)
spellingShingle Secuenciación por Nanoporos
Genética
Nanopore Sequencing
Genetics
RCCX Modules
CYP21A2
Long Read Sequencing
21-hydroxylase Deficiency
Claps, Aldana
Fernández, Cecilia
Fernandez, Franco Daniel
Macchiaroli, Natalia
Ingravidi, Marina L.
Delea, Marisol
Fernández, Cecilia
Castro, Tania
Laiseca, Julieta
Kamenetzky, Laura
Taboas, Melisa
Dain, Liliana
High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title_full High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title_fullStr High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title_full_unstemmed High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title_short High precision characterization of RCCX rearrangements in a 21-hydroxylase deficiency Latin American cohort using oxford nanopore long read sequencing
title_sort high precision characterization of rccx rearrangements in a 21 hydroxylase deficiency latin american cohort using oxford nanopore long read sequencing
topic Secuenciación por Nanoporos
Genética
Nanopore Sequencing
Genetics
RCCX Modules
CYP21A2
Long Read Sequencing
21-hydroxylase Deficiency
url http://hdl.handle.net/20.500.12123/23066
https://www.nature.com/articles/s41598-025-03799-7
https://doi.org/10.1038/s41598-025-03799-7
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