Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d

© 2024 The Authors.

Detalles Bibliográficos
Autores: Abugattas-Nuñez del Prado, Joaquin A., Ding, Yi, Sonneville, Jan de, Kolk, Kees-Jan van der, Moreno-Mateos, Miguel A., Málaga-Trillo, Edward, Spaink, Herman P.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/372326
Acceso en línea:http://hdl.handle.net/10261/372326
Access Level:acceso abierto
Palabra clave:CasRx
High throughput
Knockdown
Robot microinjector
Zebrafish
id ES_021c750ef7cbcafcfbfa9f71b9b6ce03
oai_identifier_str oai:digital.csic.es:10261/372326
network_acronym_str ES
network_name_str España
repository_id_str
spelling Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13dAbugattas-Nuñez del Prado, Joaquin A.Ding, YiSonneville, Jan deKolk, Kees-Jan van derMoreno-Mateos, Miguel A.Málaga-Trillo, EdwardSpaink, Herman P.CasRxHigh throughputKnockdownRobot microinjectorZebrafish© 2024 The Authors.In this study, the authors compared the efficiency of automated robotic and manual injection methods for the CRISPR-RfxCas13d (CasRx) system for mRNA knockdown and Cas9-mediated DNA targeting in zebrafish embryos. They targeted the no tail (TBXTA) gene as a proof-of-principle, evaluating the induced embryonic phenotypes. Both Cas9 and CasRx systems caused loss of function phenotypes for TBXTA. Cas9 protein exhibited a higher percentage of severe phenotypes compared with mRNA, while CasRx protein and mRNA showed similar efficiency. Both robotic and manual injections demonstrated comparable phenotype percentages and mortality rates. The findings highlight the potential of RNA-targeting CRISPR effectors for precise gene knockdown and endorse automated microinjection at a speed of 1.0 s per embryo as a high-throughput alternative to manual methods.Work in MA Moreno-Mateos' lab was supported by grants PID2021-127535NB-I00, CEX2020-001088-M and RyC-2017-23041 funded by MCIN/AEI/ 10.13039/501100011033 by “ERDF A way of making Europe” and by ESF Investing in your future, grant UPO1380590 funded by “ERDF A way of making Europe”, University Pablo de Olavide and Junta de Andalucía.With funding from the Spanish government through the ‘María de Maeztu Unit of Excelence’ accreditation (CEX2020-001088-M)Peer reviewedTaylor & FrancisMinisterio de Ciencia e Innovación (España)Agencia Estatal de Investigación (España)Ministerio de Ciencia, Innovación y Universidades (España)European CommissionUniversidad Pablo de OlavideJunta de AndalucíaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/372326reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127535NB-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/CEX2020-001088-MAbugattas-Nuñez del Prado, Joaquin A.; Ding, Yi; Sonneville, Jan de; Kolk, Kees-Jan van der; Moreno-Mateos, Miguel A.; Málaga-Trillo, Edward; Spaink, Herman P.; 2024; Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d [Dataset]; Taylor & Francis; https://doi.org/10.6084/m9.figshare.26068321.v1https://doi.org/10.2144/btn-2023-0062Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3723262026-05-22T06:33:51Z
dc.title.none.fl_str_mv Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
title Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
spellingShingle Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
Abugattas-Nuñez del Prado, Joaquin A.
CasRx
High throughput
Knockdown
Robot microinjector
Zebrafish
title_short Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
title_full Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
title_fullStr Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
title_full_unstemmed Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
title_sort Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
dc.creator.none.fl_str_mv Abugattas-Nuñez del Prado, Joaquin A.
Ding, Yi
Sonneville, Jan de
Kolk, Kees-Jan van der
Moreno-Mateos, Miguel A.
Málaga-Trillo, Edward
Spaink, Herman P.
author Abugattas-Nuñez del Prado, Joaquin A.
author_facet Abugattas-Nuñez del Prado, Joaquin A.
Ding, Yi
Sonneville, Jan de
Kolk, Kees-Jan van der
Moreno-Mateos, Miguel A.
Málaga-Trillo, Edward
Spaink, Herman P.
author_role author
author2 Ding, Yi
Sonneville, Jan de
Kolk, Kees-Jan van der
Moreno-Mateos, Miguel A.
Málaga-Trillo, Edward
Spaink, Herman P.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Agencia Estatal de Investigación (España)
Ministerio de Ciencia, Innovación y Universidades (España)
European Commission
Universidad Pablo de Olavide
Junta de Andalucía
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv CasRx
High throughput
Knockdown
Robot microinjector
Zebrafish
topic CasRx
High throughput
Knockdown
Robot microinjector
Zebrafish
description © 2024 The Authors.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/372326
url http://hdl.handle.net/10261/372326
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127535NB-I00
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/CEX2020-001088-M
Abugattas-Nuñez del Prado, Joaquin A.; Ding, Yi; Sonneville, Jan de; Kolk, Kees-Jan van der; Moreno-Mateos, Miguel A.; Málaga-Trillo, Edward; Spaink, Herman P.; 2024; Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d [Dataset]; Taylor & Francis; https://doi.org/10.6084/m9.figshare.26068321.v1
https://doi.org/10.2144/btn-2023-0062
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Taylor & Francis
publisher.none.fl_str_mv Taylor & Francis
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869402631876640768
score 15.812455

Similares en LA Referencia