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Use of vivo-morpholinos for gene knockdown in the postnatal shark retina

Mariña Rodríguez-Arrizabalaga, Ismael Hernández-Núñez, Eva Candal, View ORCID ProfileAntón Barreiro-Iglesias
doi: https://doi.org/10.1101/2022.07.11.499558
Mariña Rodríguez-Arrizabalaga
1Departamento de Bioloxía Funcional, CIBUS, Facultade de Bioloxía, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
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Ismael Hernández-Núñez
1Departamento de Bioloxía Funcional, CIBUS, Facultade de Bioloxía, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
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Eva Candal
1Departamento de Bioloxía Funcional, CIBUS, Facultade de Bioloxía, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
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Antón Barreiro-Iglesias
1Departamento de Bioloxía Funcional, CIBUS, Facultade de Bioloxía, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
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  • ORCID record for Antón Barreiro-Iglesias
  • For correspondence: anton.barreiro@usc.es
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Abstract

Work in the catshark Scyliorhinus canicula has shown that the evolutionary origin of postnatal neurogenesis in vertebrates is earlier than previously thought. Thus, the catshark can serve as a model of interest to understand postnatal neurogenic processes and their evolution in vertebrates. One of the best characterized neurogenic niches of the catshark CNS is found in the peripheral region of the retina. Unfortunately, the lack of genetic tools in sharks limits the possibilities to deepen in the study of genes involved in the neurogenic process. Here, we report a method for gene knockdown in the juvenile catshark retina based on the use of Vivo-Morpholinos. To establish the method, we designed Vivo-Morpholinos against the proliferation marker PCNA. We first evaluated the possible toxicity of 3 different intraocular administration regimes. After this optimization step, we show that a single intraocular injection of the PCNA Vivo-Morpholino decreases the expression of PCNA in the peripheral retina, which leads to reduced mitotic activity in this region. This method will help in deciphering the role of other genes potentially involved in postnatal neurogenesis in this model.

Summary statement In this study, we report the development of a method for the use of Vivo-Morpholinos in the postnatal shark retina, which will allow to decipher the role of different genes in retinal neurogenesis in sharks.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted July 12, 2022.
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Use of vivo-morpholinos for gene knockdown in the postnatal shark retina
Mariña Rodríguez-Arrizabalaga, Ismael Hernández-Núñez, Eva Candal, Antón Barreiro-Iglesias
bioRxiv 2022.07.11.499558; doi: https://doi.org/10.1101/2022.07.11.499558
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Use of vivo-morpholinos for gene knockdown in the postnatal shark retina
Mariña Rodríguez-Arrizabalaga, Ismael Hernández-Núñez, Eva Candal, Antón Barreiro-Iglesias
bioRxiv 2022.07.11.499558; doi: https://doi.org/10.1101/2022.07.11.499558

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