Exploring aptamers for targeted enrichment of X sperm in bovine: unraveling selective potential.
Anim Biotechnol
; 35(1): 2323592, 2024 Nov.
Article
em En
| MEDLINE
| ID: mdl-38770771
ABSTRACT
Nucleic acid aptamers have been used in the past for the development of diagnostic methods against a number of targets such as bacteria, pesticides, cancer cells etc. In the present study, six rounds of Cell-SELEX were performed on a ssDNA aptamer library against X-enriched sperm cells from Sahiwal breed cattle. Sequencing was used to examine the aptamer sequences that shown affinity for sperm carrying the X chromosome in order to find any possible X-sperm-specific sequences. Out of 35 identified sequences, 14 were selected based on bioinformatics analysis like G-Score and Mfold structures. Further validation of their specificity was done via fluorescence microscopy. The interaction of biotinylated-aptamer with sperm was also determined by visualizing the binding of streptavidin coated magnetic beads on the head region of the sperm under bright field microscopy. Finally, a real-time experiment was designed for the validation of X-sperm enrichment by synthesized aptamer sequences. Among the studied sequences, aptamer 29a exhibited a higher affinity for X sperm compared to Y sperm in a mixed population of sperm cells. By using aptamer sequence 29a, we obtained an enrichment of 70% for X chromosome bearing sperm cells.
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Base de dados:
MEDLINE
Assunto principal:
Espermatozoides
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Cromossomo X
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Aptâmeros de Nucleotídeos
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Técnica de Seleção de Aptâmeros
Limite:
Animals
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article