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A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene

Amphioxus is a promising model organism for understanding the origin and evolution of vertebrates due to its basal phylogenetic position among chordates. We here compared the mutation efficacy and mutation type of tail tips and gametes of amphioxus founders injected with Cas9 protein and six differe...

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Autores principales: Zou, Jiaqi, Wu, Xiaotong, Shi, Chenggang, Zhong, Yanhong, Zhang, Lei, Yan, Qiuning, Su, Liuru, Li, Guang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8387717/
https://www.ncbi.nlm.nih.gov/pubmed/34458263
http://dx.doi.org/10.3389/fcell.2021.702290
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author Zou, Jiaqi
Wu, Xiaotong
Shi, Chenggang
Zhong, Yanhong
Zhang, Lei
Yan, Qiuning
Su, Liuru
Li, Guang
author_facet Zou, Jiaqi
Wu, Xiaotong
Shi, Chenggang
Zhong, Yanhong
Zhang, Lei
Yan, Qiuning
Su, Liuru
Li, Guang
author_sort Zou, Jiaqi
collection PubMed
description Amphioxus is a promising model organism for understanding the origin and evolution of vertebrates due to its basal phylogenetic position among chordates. We here compared the mutation efficacy and mutation type of tail tips and gametes of amphioxus founders injected with Cas9 protein and six different sgRNAs targeting five distinct genes, and revealed a strong correlation for mutation efficacy and a mild correlation for mutation type among the two tissues. In addition, we also observed a positive relationship between gene insertions observed in tail tips and gametes of amphioxus founders injected with Tol2 transposase and two different transgenic constructs. Finally, we showed that amphioxus larvae which had their tail tips cut at the 3–4 gill-slit stage were able to recover within 6 days and developed a normal number of gonads at the adult stage, and that F0 larvae carry similar mutation efficacy and type in the posterior end to that in the tail tips after their metamorphosis. Together, these findings suggest a great potential for obtaining valid amphioxus founders with desired mutations and transgenes at as early as the early larval stage, which will certainly speed up the generation of amphioxus mutants and transgenes and make it more cost- and labor-effective.
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spelling pubmed-83877172021-08-27 A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene Zou, Jiaqi Wu, Xiaotong Shi, Chenggang Zhong, Yanhong Zhang, Lei Yan, Qiuning Su, Liuru Li, Guang Front Cell Dev Biol Cell and Developmental Biology Amphioxus is a promising model organism for understanding the origin and evolution of vertebrates due to its basal phylogenetic position among chordates. We here compared the mutation efficacy and mutation type of tail tips and gametes of amphioxus founders injected with Cas9 protein and six different sgRNAs targeting five distinct genes, and revealed a strong correlation for mutation efficacy and a mild correlation for mutation type among the two tissues. In addition, we also observed a positive relationship between gene insertions observed in tail tips and gametes of amphioxus founders injected with Tol2 transposase and two different transgenic constructs. Finally, we showed that amphioxus larvae which had their tail tips cut at the 3–4 gill-slit stage were able to recover within 6 days and developed a normal number of gonads at the adult stage, and that F0 larvae carry similar mutation efficacy and type in the posterior end to that in the tail tips after their metamorphosis. Together, these findings suggest a great potential for obtaining valid amphioxus founders with desired mutations and transgenes at as early as the early larval stage, which will certainly speed up the generation of amphioxus mutants and transgenes and make it more cost- and labor-effective. Frontiers Media S.A. 2021-08-12 /pmc/articles/PMC8387717/ /pubmed/34458263 http://dx.doi.org/10.3389/fcell.2021.702290 Text en Copyright © 2021 Zou, Wu, Shi, Zhong, Zhang, Yan, Su and Li. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Zou, Jiaqi
Wu, Xiaotong
Shi, Chenggang
Zhong, Yanhong
Zhang, Lei
Yan, Qiuning
Su, Liuru
Li, Guang
A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title_full A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title_fullStr A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title_full_unstemmed A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title_short A Potential Method for Rapid Screening of Amphioxus Founder Harboring Germline Mutation and Transgene
title_sort potential method for rapid screening of amphioxus founder harboring germline mutation and transgene
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8387717/
https://www.ncbi.nlm.nih.gov/pubmed/34458263
http://dx.doi.org/10.3389/fcell.2021.702290
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