A single blastomere sexing of caprine embryos by simultaneous amplification of sex chromosome-specific sequence of SRY and amelogenin genes

Malik, Hruda Nanda; Singhal, Dinesh Kumar; Mukherjee, Ayan; Bara, Nisha; Kumar, Sudarshan; Saugandhika, Shrabani; Mohanty, Ashok Kumar; Kaushik, Jai K.; Bag, Sadhan; Das, Bikash; Bhanja, S.K.; Malakar, Dhruba

doi:10.1016/j.livsci.2013.05.030

Cited by 5 publications

(4 citation statements)

References 28 publications

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“…Tests which amplify homologous X‐ and Y‐specific genes with the same pair of primers already include the internal control. Nevertheless, an additional primer pair for a Y‐specific gene (mostly SRY ) can still be included when developing a method, in order to corroborate the results (Lindsay & Belant, ; Malik et al, 2013; Morin et al, ).…”

Section: Resultsmentioning

confidence: 99%

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Molecular sexing assays in 114 mammalian species: In silico sequence reanalysis and a unified graphical visualization of diagnostic tests

Strah

Kunej

2019

Ecology and Evolution

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Molecular‐based methods for identifying sex in mammals have a wide range of applications, from embryo manipulation to ecological studies. Various sex‐specific or homologous genes can be used for this purpose, PCR amplification being a common method. Over the years, the number of reported tests and the range of tested species have increased greatly. The aim of the present analysis was to retrieve PCR‐based sexing assays for a range of mammalian species, gathering the gene sequences from either the articles or online databases, and visualize the molecular design in a uniform manner. For nucleotide alignment and diagnostic test visualization, the following genomic databases and tools were used: NCBI, Ensembl Nucleotide BLAST, ClustalW2, and NEBcutter V2.0. In the 45 gathered articles, 59 different diagnostic tests based on eight different PCR‐based methods were developed for 114 mammalian species. Most commonly used genes for the analysis were ZFX , ZFY , AMELX , and AMELY . The tests were most commonly based on sex‐specific insertions and deletions (SSIndels) and sex‐specific sequence polymorphisms (SSSP). This review provides an overview of PCR‐based sexing methods developed for mammals. This information will facilitate more efficient development of novel molecular sexing assays and reuse of previously developed tests. Development of many novel and improvement of previously developed tests is also expected with the rapid increase in the quantity and quality of available genetic information.

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Section: Resultsmentioning

confidence: 99%

“…Nevertheless, an additional primer pair for a Y-specific gene (mostly SRY) can still be included when developing a method, in order to corroborate the results (Lindsay & Belant, 2007;Malik et al, 2013;Morin et al, 2005).…”

Section: Internal Amplification Controlsmentioning

confidence: 99%

Molecular sexing assays in 114 mammalian species: In silico sequence reanalysis and a unified graphical visualization of diagnostic tests

Strah

Kunej

2019

Ecology and Evolution

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“…The advantages include an approach that is extremely responsive, accurate, dependable, and efficient, and also a consistent pregnancy rate. Considerable many embryos can be sexed using this technique (Malik et al, 2013). Embryo biopsy can be used to perform multiplex genotyping on bovine embryos as well as genetic tests for inherited diseases (Peippo et al, 2007).…”

Section: ) Polymerase Chain Reactionmentioning

confidence: 99%

Embryo sexing methods in bovine and its application in animal breed

Bora

2022

J Anim Reprod Biotechnol

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The ability to determine the gender of bovine embryos before the transfer is useful in livestock management, especially in dairy production, where female calves are favored. The milk production and meat are the result of both female and male cattle that benefits the dairy and beef industries (Sachan et al., 2020). Pre-implantation sexing of embryos also assists in the effectiveness of embryo transfers. In farm animals, there are two methods for sexing bovine embryos: invasive and non-invasive.The non-invasive treatment is considered the best since it preserves the embryo's autonomy. The invasive method does not protect the autonomy of an embryo and is likely to affect the embryo's chances of successful transfer. The milk quality and meat of both female and male cattle

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“…Genotyping of putative eScs. The sexing or genotyping of ESCs colonies was performed as described by earlier reports 49 . The ESCs colonies were manually sliced into very small pieces and cultured in a hanging drop (25 µl) of EB specific culture medium (DMEM supplemented with 5% knock out serum replacement, 0.5% (100×) non-essential amino acid and 1% essential amino acid) at 37 °C in 5% air for 3 days to develop into embryoid bodies.…”

Section: Animal Ethics All Experiments Were Conducted As Per the Guimentioning

confidence: 99%

Derivation of oocyte-like cells from putative embryonic stem cells and parthenogenetically activated into blastocysts in goat

Malik

Singhal

Saini

et al. 2020

Sci Rep

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Germ cells are responsible for the propagation of live animals from generation to generation, but to surprise, a steep increase in infertile problems among livestock poses great threat for economic development of human race. An alternative and robust approach is essential to combat these ailments. Here, we demonstrate that goat putative embryonic stem cells (eScs) were successfully in vitro differentiated into primordial germ cells and oocyte-like cells using bone morphogenetic protein-4 (BMP-4) and trans-retinoic acid (RA). Oocyte-like cells having distinct zonapellucida recruited adjacent somatic cells in differentiating culture to form cumulus-oocyte complexes (COCs). The putative COCs were found to express the zonapellucida specific (ZP1 and ZP2) and oocyte-specific markers. Primordial germ cell-specific markers VASA, DAZL, STELLA, and PUM1 were detected at protein and mRNA level. In addition to that, the surface architecture of these putative COCs was thoroughly visualized by the scanning electron microscope. the putative cocs were further parthenogenetically activated to develop into healthy morula, blastocysts and hatched blastocyst stage like embryos. Our findings may contribute to the fundamental understanding of mammalian germ cell biology and may provide clinical insights regarding infertility ailments. Germ cells hold an idiosyncratic and crucial position in the biorhythms of animals as they pass the genome onto the succeeding generation 1. In mammals the primordial germ cells (PGCs) are derived from proximal epiblast of a developing embryo 2. Once developed, PGCs drifted through dorsal mesentery of hindgut and invade the genital ridge of developing fetus in mouse 3. Large size, low nucleo-cytoplasm ratio, distinct nuclear borders, and granular nuclear chromatin are the important characters of PGCs 4. Embryonic stem cells (ESCs) are emanated from blastocyst stage embryos in mammals with unlimited proliferative capacity and multi lineages differentiation ability under suitable culture onditions 5-8. These cells hold a wide prospective to bestead not only as in vitro model system to extrapolate the mechanism of gametogenesis but also a cell based method of inducing mature gametes. Derivation of mature germ cells from ESCs was obtained in two ways: first, spontaneous differentiation of adherent cultures after discarding feeder layers and leukemia inhibitory factor 9-11 and second, embryoid body-mediated germ cell production 12-17. Moreover several aspects in germ cells genesis from ESCs i.e. lack of reproducibility and validity of ESCs-derived germ cell identity were not well established 18-20. The definitive markers of female germ cells in both XX and XY cell lines is expressed during ESCs development 12,13. The prevailing hypothesis that regardless of the sex of germ cells, they are inherently destined to undergo meiosis and develop as oocytes, unless otherwise prevented by factors inhabiting meiosis from doing so 1. Although it has been shown to be possible to distinguish cells resembling germ cells f...

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A single blastomere sexing of caprine embryos by simultaneous amplification of sex chromosome-specific sequence of SRY and amelogenin genes

Cited by 5 publications

References 28 publications

Molecular sexing assays in 114 mammalian species: In silico sequence reanalysis and a unified graphical visualization of diagnostic tests

Molecular sexing assays in 114 mammalian species: In silico sequence reanalysis and a unified graphical visualization of diagnostic tests

Embryo sexing methods in bovine and its application in animal breed

Derivation of oocyte-like cells from putative embryonic stem cells and parthenogenetically activated into blastocysts in goat

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