[MARMAM] New publication on a mammalian mRNA sex determination method

jake linsky jmjlinsky at gmail.com
Wed Aug 24 19:09:02 PDT 2022


Dear Colleagues,

   My co-authors and I are happy to share our new paper "A mammalian
messenger RNA sex determination method from humpback whale (Megaptera
novaeangliae) blubber biopsies" in The Royal Society Open Science. Our
paper presents a functional blubber mRNA extraction protocol and a method
of determining sex using the extracted mRNA, to help streamline
expression-based health studies.

A mammalian messenger RNA sex determination method from humpback whale
(Megaptera novaeangliae) blubber biopsies
Jacob M. J. Linsky, Rebecca A. Dunlop, Michael J. Noad and Lee A. McMichael
Published:24 August 2022 https://doi.org/10.1098/rsos.220556

Abstract
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.

A mammalian messenger RNA sex determination method from humpback whale
(Megaptera novaeangliae) blubber biopsies | Royal Society Open Science
(royalsocietypublishing.org)
<https://royalsocietypublishing.org/doi/10.1098/rsos.220556>

regards,
-Jake

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