A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies
The large size of free-ranging mysticetes, such as humpback whales (Megaptera novaeangliae), make capture and release health assessments unfeasible for conservation research. However, individual energetic condition or reproductive health may be assessed from the gene expression of remotely biopsied...
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ftpubmed:oai:pubmedcentral.nih.gov:9399696 2023-05-15T16:36:05+02:00 A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies Linsky, Jacob M. J. Dunlop, Rebecca A. Noad, Michael J. McMichael, Lee A. 2022-08-24 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399696/ https://doi.org/10.1098/rsos.220556 en eng The Royal Society http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399696/ http://dx.doi.org/10.1098/rsos.220556 © 2022 The Authors. https://creativecommons.org/licenses/by/4.0/Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, provided the original author and source are credited. CC-BY R Soc Open Sci Genetics and Genomics Text 2022 ftpubmed https://doi.org/10.1098/rsos.220556 2022-08-28T01:09:53Z The large size of free-ranging mysticetes, such as humpback whales (Megaptera novaeangliae), make capture and release health assessments unfeasible for conservation research. However, individual energetic condition or reproductive health may be assessed from the gene expression of remotely biopsied tissue. To do this, researchers must reliably extract RNA and interpret gene expression measurements within the context of an individual's sex. Here, we outline an RNA extraction protocol from blubber tissue and describe a novel mammalian RNA sex determination method. Our method consists of a duplex reverse transcription-quantitative (real-time) polymerase chain reaction (RT-qPCR) with primer sets for a control gene (ACTB) and the X-chromosome inactivation gene (XIST). Products of each RT-qPCR had distinct melting temperature profiles based on the presence (female) or absence (male) of the XIST transcript. Using high-resolution melt analysis, reactions were sorted into one of two clusters (male/female) based on their melting profiles. We validated the XIST method by comparing results with a standard DNA-based method. With adequate quantities of RNA (minimum of approx. 9 ng µl(−1)), the XIST sex determination method shows 100% agreement with traditional DNA sex determination. Using the XIST method, future cetacean health studies can interpret gene expression within the context of an individual's sex, all from a single extraction. Text Humpback Whale Megaptera novaeangliae PubMed Central (PMC) Royal Society Open Science 9 8 |
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ftpubmed |
language |
English |
topic |
Genetics and Genomics |
spellingShingle |
Genetics and Genomics Linsky, Jacob M. J. Dunlop, Rebecca A. Noad, Michael J. McMichael, Lee A. A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
topic_facet |
Genetics and Genomics |
description |
The large size of free-ranging mysticetes, such as humpback whales (Megaptera novaeangliae), make capture and release health assessments unfeasible for conservation research. However, individual energetic condition or reproductive health may be assessed from the gene expression of remotely biopsied tissue. To do this, researchers must reliably extract RNA and interpret gene expression measurements within the context of an individual's sex. Here, we outline an RNA extraction protocol from blubber tissue and describe a novel mammalian RNA sex determination method. Our method consists of a duplex reverse transcription-quantitative (real-time) polymerase chain reaction (RT-qPCR) with primer sets for a control gene (ACTB) and the X-chromosome inactivation gene (XIST). Products of each RT-qPCR had distinct melting temperature profiles based on the presence (female) or absence (male) of the XIST transcript. Using high-resolution melt analysis, reactions were sorted into one of two clusters (male/female) based on their melting profiles. We validated the XIST method by comparing results with a standard DNA-based method. With adequate quantities of RNA (minimum of approx. 9 ng µl(−1)), the XIST sex determination method shows 100% agreement with traditional DNA sex determination. Using the XIST method, future cetacean health studies can interpret gene expression within the context of an individual's sex, all from a single extraction. |
format |
Text |
author |
Linsky, Jacob M. J. Dunlop, Rebecca A. Noad, Michael J. McMichael, Lee A. |
author_facet |
Linsky, Jacob M. J. Dunlop, Rebecca A. Noad, Michael J. McMichael, Lee A. |
author_sort |
Linsky, Jacob M. J. |
title |
A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
title_short |
A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
title_full |
A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
title_fullStr |
A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
title_full_unstemmed |
A mammalian messenger RNA sex determination method from humpback whale (Megaptera novaeangliae) blubber biopsies |
title_sort |
mammalian messenger rna sex determination method from humpback whale (megaptera novaeangliae) blubber biopsies |
publisher |
The Royal Society |
publishDate |
2022 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399696/ https://doi.org/10.1098/rsos.220556 |
genre |
Humpback Whale Megaptera novaeangliae |
genre_facet |
Humpback Whale Megaptera novaeangliae |
op_source |
R Soc Open Sci |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399696/ http://dx.doi.org/10.1098/rsos.220556 |
op_rights |
© 2022 The Authors. https://creativecommons.org/licenses/by/4.0/Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, provided the original author and source are credited. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1098/rsos.220556 |
container_title |
Royal Society Open Science |
container_volume |
9 |
container_issue |
8 |
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1766026393905266688 |