Selection of sex-specific aptamer probes to sperm

Colley, Adam J.; Buhr, M.M.; Golovan, Serguei P.

doi:10.1016/j.theriogenology.2008.06.039

Cited by 2 publications

(3 citation statements)

References 134 publications

(195 reference statements)

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“…Given the size of bovine genome to be 3 to 4 GB and a motif of approximately 10 nucleotides long is expected to occur at the rate of 43 times in the genome by chance. The genome of spermatozoa is highly condensed [ 33 ] and hence the genomic DNA may also be inaccessible for the oligonucleotides to bind in the conditions applied and hence lowering the possibility of enriching amplicons to sperm DNA [ 21 ]. In addition, the 27-nucleotide primer binding site on BLASTN analysis did not reveal any perfect or near perfect similarity to sequences in the bovine genome.…”

Section: Discussionmentioning

confidence: 99%

“…Aptamers have been generated for various biomedical application including therapeutics [11,12,13,14], aptasensors [15], biosensors [16,17], diagnostic [18,19], and imaging systems [20]. Efforts have been made to generate X-and Y-sex-specific oligonucleotide from a large randomised library to generate oligonucleotides that selectively bind to boar X-or Y-spermatozoa [21]. Due to above inherent advantages, as a first step we choose to use a random oligonucleotide library, perform SELEX rounds with bovine spermatozoa and identify oligonucleotides that exhibit binding to bovine spermatozoa.…”

Section: A C C E P T E Dmentioning

confidence: 99%

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Generation of single stranded DNA with selective affinity to bovine spermatozoa

et al. 2021

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Objective: This study was conducted to generate single stranded DNA oligonucleotides with selective affinity to bovine spermatozoa, assess its binding potential and explore its potential utility in trapping spermatozoa from suspensions.Methods: A combinatorial library of 94 mer long oligonucleotide was used for systematic evolution of ligands by exponential enrichment (SELEX) with bovine spermatozoa. The amplicons from sixth and seventh rounds of SELEX were sequenced, and the reads were clustered employing cluster database at high identity with tolerance (CD-HIT) and FASTAptamer. The enriched nucleotides were predicted for secondary structures by Mfold, motifs by Multiple Em for Motif Elicitation and 5’ labelled with biotin/6-FAM to determine the binding potential and binding pattern.Results: We generated 14.1 and 17.7 million reads from sixth and seventh rounds of SELEX respectively to bovine spermatozoa. The CD-HIT clustered 78,098 and 21,196 reads in the top ten clusters and FASTAptamer identified 2,195 and 4,405 unique sequences in the top three clusters from the sixth and seventh rounds, respectively. The identified oligonucleotides formed secondary structures with delta G values between –1.17 to –26.18 kcal/mol indicating varied stability. Confocal imaging with the oligonucleotides from the seventh round revealed different patterns of binding to bovine spermatozoa (fluorescence of the whole head, spot of fluorescence in head and mid- piece and tail). Use of a 5’-biotin tagged oligonucleotide from the sixth round at 100 pmol with 4×106 spermatozoa could trap almost 80% from the suspension.Conclusion: The binding patterns and ability of the identified oligonucleotides confirms successful optimization of the SELEX process and generation of aptamers to bovine spermatozoa. These oligonucleotides provide a quick approach for selective capture of spermatozoa from complex samples. Future SELEX rounds with X- or Y- enriched sperm suspension will be used to generate oligonucleotides that bind to spermatozoa of a specific sex type.

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Section: Discussionmentioning

confidence: 99%

Section: A C C E P T E Dmentioning

confidence: 99%

Generation of single stranded DNA with selective affinity to bovine spermatozoa

et al. 2021

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“…Based on these tests, aptamers can be easily chosen for sperm features, and similar methods can be useful for further work in the use of antibodies. Besides, X and Y-sperm can show a sex-unique marker that can be resolved using aptamer probes (Colley 2008).…”

Section: Recent Advances In Sperm Sexingmentioning

confidence: 99%

Use of sexed semen for genetic improvement of indigenous dairy cattle and buffaloes productivity

Yata

Singh

Kumar

et al. 2022

Indian J of Anim Sci

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The productivity of dairy animals in India needs to be improved to feed the growing population in the decades ahead. Sperm sexing offer a promising breeding strategy to meet the increased demand for food production and nutritional protection. Application of sexed sperm in the breeding of indigenous cattle and buffalo would help to protect endangered species, and it also helps to improve the productivity of indigenous milch animals which can survive for long starvation and well adopted by Indian climatic conditions with disease resistance. Â The productivity of crossbreed milch animals also required to meet the food demand associated with population growth. This review discusses the sustainability of Indian dairy animals and provides sperm sexing strategies to boost dairy animal production in India.

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Selection of sex-specific aptamer probes to sperm

Cited by 2 publications

References 134 publications

Generation of single stranded DNA with selective affinity to bovine spermatozoa

Generation of single stranded DNA with selective affinity to bovine spermatozoa

Use of sexed semen for genetic improvement of indigenous dairy cattle and buffaloes productivity

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