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Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development

BACKGROUND: The Natterin protein family was first discovered in the venom of the medically significant fish Thalassophryne nattereri, and over the last decade natterin-like genes have been identified in various organisms, notably performing immune-related functions. Previous findings support natteri...

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Autores principales: Seni-Silva, Ana Carolina, Maleski, Adolfo Luis Almeida, Souza, Milena Marcolino, Falcao, Maria Alice Pimentel, Disner, Geonildo Rodrigo, Lopes-Ferreira, Monica, Lima, Carla
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8840632/
https://www.ncbi.nlm.nih.gov/pubmed/35151271
http://dx.doi.org/10.1186/s12864-022-08369-z
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author Seni-Silva, Ana Carolina
Maleski, Adolfo Luis Almeida
Souza, Milena Marcolino
Falcao, Maria Alice Pimentel
Disner, Geonildo Rodrigo
Lopes-Ferreira, Monica
Lima, Carla
author_facet Seni-Silva, Ana Carolina
Maleski, Adolfo Luis Almeida
Souza, Milena Marcolino
Falcao, Maria Alice Pimentel
Disner, Geonildo Rodrigo
Lopes-Ferreira, Monica
Lima, Carla
author_sort Seni-Silva, Ana Carolina
collection PubMed
description BACKGROUND: The Natterin protein family was first discovered in the venom of the medically significant fish Thalassophryne nattereri, and over the last decade natterin-like genes have been identified in various organisms, notably performing immune-related functions. Previous findings support natterin-like genes as effector defense molecules able to activate multiprotein complexes driving the host innate immune response, notably due to the pore-forming function of the aerolysin superfamily members. Herein, employing a combination of the CRISPR/Cas9 depletion system, phenotype-based screening, and morphometric methods, we evaluated the role of one family member, LOC795232, in the embryonic development of zebrafish since it might be implicated in multiple roles and characterization of the null mutant is central for analysis of gene activity. RESULTS: Multiple sequence alignment revealed that the candidate natterin-like has the highest similarity to zebrafish aep1, a putative and better characterized fish-specific defense molecule from the same family. Compared to other species, zebrafish have many natterin-like copies. Whole-mount in situ hybridization confirmed the knockout and mutant embryos exhibited epiboly delay, growth retardation, yolk sac and heart edema, absent or diminished swim bladder, spinal defects, small eyes and head, heart dysfunction, and behavioral impairment. As previously demonstrated, ribonucleoproteins composed of Cas9 and duplex guide RNAs are effective at inducing mutations in the F0 zebrafish. CONCLUSIONS: The considerably high natterin-like copies in zebrafish compared to other species might be due to the teleost-specific whole genome duplication and followed by subfunctionalization or neofunctionalization. In the present work, we described some of the natterin-like features in the zebrafish development and infer that natterin-like proteins potentially contribute to the embryonary development and immune response. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-022-08369-z.
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spelling pubmed-88406322022-02-16 Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development Seni-Silva, Ana Carolina Maleski, Adolfo Luis Almeida Souza, Milena Marcolino Falcao, Maria Alice Pimentel Disner, Geonildo Rodrigo Lopes-Ferreira, Monica Lima, Carla BMC Genomics Research BACKGROUND: The Natterin protein family was first discovered in the venom of the medically significant fish Thalassophryne nattereri, and over the last decade natterin-like genes have been identified in various organisms, notably performing immune-related functions. Previous findings support natterin-like genes as effector defense molecules able to activate multiprotein complexes driving the host innate immune response, notably due to the pore-forming function of the aerolysin superfamily members. Herein, employing a combination of the CRISPR/Cas9 depletion system, phenotype-based screening, and morphometric methods, we evaluated the role of one family member, LOC795232, in the embryonic development of zebrafish since it might be implicated in multiple roles and characterization of the null mutant is central for analysis of gene activity. RESULTS: Multiple sequence alignment revealed that the candidate natterin-like has the highest similarity to zebrafish aep1, a putative and better characterized fish-specific defense molecule from the same family. Compared to other species, zebrafish have many natterin-like copies. Whole-mount in situ hybridization confirmed the knockout and mutant embryos exhibited epiboly delay, growth retardation, yolk sac and heart edema, absent or diminished swim bladder, spinal defects, small eyes and head, heart dysfunction, and behavioral impairment. As previously demonstrated, ribonucleoproteins composed of Cas9 and duplex guide RNAs are effective at inducing mutations in the F0 zebrafish. CONCLUSIONS: The considerably high natterin-like copies in zebrafish compared to other species might be due to the teleost-specific whole genome duplication and followed by subfunctionalization or neofunctionalization. In the present work, we described some of the natterin-like features in the zebrafish development and infer that natterin-like proteins potentially contribute to the embryonary development and immune response. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-022-08369-z. BioMed Central 2022-02-12 /pmc/articles/PMC8840632/ /pubmed/35151271 http://dx.doi.org/10.1186/s12864-022-08369-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Seni-Silva, Ana Carolina
Maleski, Adolfo Luis Almeida
Souza, Milena Marcolino
Falcao, Maria Alice Pimentel
Disner, Geonildo Rodrigo
Lopes-Ferreira, Monica
Lima, Carla
Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title_full Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title_fullStr Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title_full_unstemmed Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title_short Natterin-like depletion by CRISPR/Cas9 impairs zebrafish (Danio rerio) embryonic development
title_sort natterin-like depletion by crispr/cas9 impairs zebrafish (danio rerio) embryonic development
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8840632/
https://www.ncbi.nlm.nih.gov/pubmed/35151271
http://dx.doi.org/10.1186/s12864-022-08369-z
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