Materials and methods for sperm sex selection

A sperm and semen technology, applied in chemical instruments and methods, material testing products, medical raw materials derived from mammals, etc., can solve problems such as difficult analysis, unsuccessful discovery, limited dynamic range, etc., and achieve high sensitivity and high specificity. Effect

Inactive Publication Date: 2010-09-01
ANDROGENIX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the analytical approach is still hampered by two major issues: first, the most difficult group of proteins to analyze using this system are membrane components, such as integrins, due to solubility issues; second, detection by mass spectrometry is limited by the dynamic range
This limited dynamic range translates into reduced sensitivity for detecting low-abundance molecules if other high-abundance species are present
[0008] Methods described so far have not been successful in identifying antigens specific to X or Y-bearing sperm, suggesting no and / or minor differences in terms of nature and / or abundance

Method used

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  • Materials and methods for sperm sex selection
  • Materials and methods for sperm sex selection
  • Materials and methods for sperm sex selection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Identification of candidate genes by bioinformatics

[0078] The publicly available bovine genome (available at the Ensembl site; originally published August 14, 2006; updated version published February 2007) and the publicly available human genome were used in a genome-based approach to identify Differences in the surface of sexed spermatozoa. Specifically, candidate genes were selected using the Ensembl Biomart tool (available on the Ensembl site) and the following strategy:

[0079] 1) Using Biomart to identify bovine orthologs of human X-chromosome genes, which have transmembrane domains, and check by manual analysis;

[0080] 2) Identify genes that are present on the X chromosome and have a transmembrane domain in the bovine genome by Biomart, and check by manual analysis (unsequenced bovine Y chromosome); and

[0081] 3) Bovine orthologs of human Y chromosome genes with transmembrane domains were identified using Biomart and checked by manual analysis (contain...

Embodiment 2

[0084] Prioritize candidate genes based on expression levels

[0085] Each candidate gene identified in Example 1 was checked for splice variants and, if so, exons common to all transcripts were selected. If no suitable exons were present, exons unique to each transcript were selected for primer design. Exons were used for primer design, rather than skipping introns, so that all primers were tested on genomic DNA. Control primers were also designed to ensure that there was no genomic DNA in the cDNA. Primers were designed for real-time PCR using Primer3 software (available online from SourceForge) and a product size of 80-150 bp. All primers were checked using Blast software to verify that they could not prime elsewhere in the genome (i.e. at least the 3' terminal bases of the primers could not be matched). The designed primers were then used in reverse transcription PCR studies to analyze the expression of candidate genes in bovine testis tissue cDNA and bovine genomic D...

Embodiment 3

[0096] Prioritization of candidate genes based on subcellular localization

[0097] The low expression levels of candidate genes in round spermatids suggest that candidates should be given lower priority if they are present only on non-cell surface membranes. The reason for this is that the candidate is very difficult to detect as it is already under-expressed, which in turn is only present in a low percentage of the protein on the surface. Candidate genes or their orthologs in other species are therefore tested to determine the subcellular location of the gene product. If evidence is obtained that the protein is on a membrane system other than the cell surface, then that candidate will be given lower priority. This data was combined with round spermatid expression data to generate 4 gene classes with different priorities, with class I being the highest priority. Each of the Class I bovine candidate genes identified in Table 4 below has the following characteristics:

[009...

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Abstract

Materials and methods for the separation of X- and Y-chromosome bearing sperm, for example in a semen sample, are provided. The methods involve contacting the semen sample with a binding agent, such as an antibody, that specifically binds to an antigen that is specific for an X- or Y-chromosome. Kits for use in the methods are also provided.

Description

field of invention [0001] The present application relates to methods for identifying sperm carrying an X chromosome or a Y chromosome. More specifically, the application relates to sex-specific antigens and their use in such methods. Background of the invention [0002] The ability to identify and select male and female sperm is of great value to the livestock industry, where there is an established artificial insemination market valued at more than $2 billion per year in the Organization for Economic Co-operation and Development (OECD) . This is especially important for the dairy industry, where the majority of dairy farmers in important OECD markets impregnate their cows through artificial insemination. Sexed semen offers farmers the opportunity to increase productivity and increase income. For example, application of sexed semen has a significant effect on reducing and / or eliminating minimal returns in heifers (compared to heifers). [0003] Genetic improvements leadi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53A61K35/52A61K39/395C07K16/28
CPCC07K16/2866C07K16/2803C07K16/2896C12N5/0612A61K35/52
Inventor K·赫森S·雷夫利奇
Owner ANDROGENIX
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