Sexing of cattle embryos using RNA-sequencing data or polymerase chain reaction based on a complete sequence of cattle chromosome Y

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dc.contributor.authorNix, Jada Lindsayen
dc.contributor.authorSchettini, Gustavo Pimentaen
dc.contributor.authorBiase, Fernando Henriqueen
dc.date.accessioned2023-09-12T14:31:15Zen
dc.date.available2023-09-12T14:31:15Zen
dc.date.issued2023-04en
dc.description.abstractWhen necessary, RNA-sequencing data or polymerase chain reaction (PCR) assays can be used to determine the presence of the chromosome Y (ChrY) in samples. This information allows for biological variation due to sexual dimorphism to be studied. A prime example is when researchers conduct RNA-sequencing of single embryos, or conceptuses, prior to the development of gonads. A recent publication of a complete sequence of the ChrY has removed limitations for the development of these procedures in cattle, otherwise imposed by the absence of a ChrY in the reference genome. Using the sequence of the cattle ChrY and transcriptome data, we conducted a systematic search for genes in the ChrY that are exclusively expressed in male tissues. The genes ENSBIXG00000029763, ENSBIXG00000029774, ENSBIXG00000029788, and ENSBIXG00000029892 were consistently expressed across male tissues and lowly expressed or absent in female samples. We observed that the cumulative values of counts per million were 2688-fold greater in males than the equivalent values in female samples. Thus, we deemed these genes suitable for the sexing of samples using RNA-sequencing data. We successfully used this set of genes to infer the sex of 22 cattle blastocysts (8 females and 14 males). Additionally, the completed sequence of the cattle ChrY has segments in the male-specific region that are not repeated. We designed a pair of oligonucleotides that targets one of these non-repeated regions in the male-specific sequence of the ChrY. Using this pair of oligonucleotides, in a multiplexed PCR assay with oligonucleotides that anneal to an autosome chromosome, we accurately identified the sex of cattle blastocysts. We developed efficient procedures for the sexing of samples in cattle using either transcriptome data or their DNA. The procedures using RNA-sequencing will greatly benefit researchers who work with samples limited in cell numbers which are only sufficient to produce transcriptome data. The oligonucleotides used for the accurate sexing of samples using PCR are transferable to other cattle tissue samples.en
dc.description.notesThis project was supported by Agriculture and Food Research Initiative Competitive Grant No. 2018-67015-31936 from the USDA National Institute of Food and Agriculture. The funding bodies had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.en
dc.description.sponsorshipAgriculture and Food Research Initiative Competitive [2018-67015-31936]; USDA National Institute of Food and Agricultureen
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3389/fgene.2023.1038291en
dc.identifier.eissn1664-8021en
dc.identifier.other1038291en
dc.identifier.pmid37077537en
dc.identifier.urihttp://hdl.handle.net/10919/116266en
dc.identifier.volume14en
dc.language.isoenen
dc.publisherFrontiersen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectsexingen
dc.subjectRNA-sequencingen
dc.subjectblastocysten
dc.subjectembryosen
dc.subjectPCRen
dc.titleSexing of cattle embryos using RNA-sequencing data or polymerase chain reaction based on a complete sequence of cattle chromosome Yen
dc.title.serialFrontiers in Geneticsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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