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Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus
Sea urchins are premier model organisms for the study of early development. However, the lengthy generation times of commonly used species have precluded application of stable genetic approaches. Here, we use the painted sea urchin Lytechinus pictus to address this limitation and to generate a homoz...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9245184/ https://www.ncbi.nlm.nih.gov/pubmed/35666622 http://dx.doi.org/10.1242/dev.200644 |
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author | Vyas, Himanshu Schrankel, Catherine S. Espinoza, Jose A. Mitchell, Kasey L. Nesbit, Katherine T. Jackson, Elliot Chang, Nathan Lee, Yoon Warner, Jacob Reitzel, Adam Lyons, Deirdre C. Hamdoun, Amro |
author_facet | Vyas, Himanshu Schrankel, Catherine S. Espinoza, Jose A. Mitchell, Kasey L. Nesbit, Katherine T. Jackson, Elliot Chang, Nathan Lee, Yoon Warner, Jacob Reitzel, Adam Lyons, Deirdre C. Hamdoun, Amro |
author_sort | Vyas, Himanshu |
collection | PubMed |
description | Sea urchins are premier model organisms for the study of early development. However, the lengthy generation times of commonly used species have precluded application of stable genetic approaches. Here, we use the painted sea urchin Lytechinus pictus to address this limitation and to generate a homozygous mutant sea urchin line. L. pictus has one of the shortest generation times of any currently used sea urchin. We leveraged this advantage to generate a knockout mutant of the sea urchin homolog of the drug transporter ABCB1, a major player in xenobiotic disposition for all animals. Using CRISPR/Cas9, we generated large fragment deletions of ABCB1 and used these readily detected deletions to rapidly genotype and breed mutant animals to homozygosity in the F(2) generation. The knockout larvae are produced according to expected Mendelian distribution, exhibit reduced xenobiotic efflux activity and can be grown to maturity. This study represents a major step towards more sophisticated genetic manipulation of the sea urchin and the establishment of reproducible sea urchin animal resources. |
format | Online Article Text |
id | pubmed-9245184 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-92451842022-07-07 Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus Vyas, Himanshu Schrankel, Catherine S. Espinoza, Jose A. Mitchell, Kasey L. Nesbit, Katherine T. Jackson, Elliot Chang, Nathan Lee, Yoon Warner, Jacob Reitzel, Adam Lyons, Deirdre C. Hamdoun, Amro Development Techniques and Resources Sea urchins are premier model organisms for the study of early development. However, the lengthy generation times of commonly used species have precluded application of stable genetic approaches. Here, we use the painted sea urchin Lytechinus pictus to address this limitation and to generate a homozygous mutant sea urchin line. L. pictus has one of the shortest generation times of any currently used sea urchin. We leveraged this advantage to generate a knockout mutant of the sea urchin homolog of the drug transporter ABCB1, a major player in xenobiotic disposition for all animals. Using CRISPR/Cas9, we generated large fragment deletions of ABCB1 and used these readily detected deletions to rapidly genotype and breed mutant animals to homozygosity in the F(2) generation. The knockout larvae are produced according to expected Mendelian distribution, exhibit reduced xenobiotic efflux activity and can be grown to maturity. This study represents a major step towards more sophisticated genetic manipulation of the sea urchin and the establishment of reproducible sea urchin animal resources. The Company of Biologists Ltd 2022-06-06 /pmc/articles/PMC9245184/ /pubmed/35666622 http://dx.doi.org/10.1242/dev.200644 Text en © 2022. Published by The Company of Biologists Ltd https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Techniques and Resources Vyas, Himanshu Schrankel, Catherine S. Espinoza, Jose A. Mitchell, Kasey L. Nesbit, Katherine T. Jackson, Elliot Chang, Nathan Lee, Yoon Warner, Jacob Reitzel, Adam Lyons, Deirdre C. Hamdoun, Amro Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title | Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title_full | Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title_fullStr | Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title_full_unstemmed | Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title_short | Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus |
title_sort | generation of a homozygous mutant drug transporter (abcb1) knockout line in the sea urchin lytechinus pictus |
topic | Techniques and Resources |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9245184/ https://www.ncbi.nlm.nih.gov/pubmed/35666622 http://dx.doi.org/10.1242/dev.200644 |
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