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Sperm cell separation methods and compositions containing aptamers or nucleic acid sequences for use therein

一种核酸序列、核苷酸序列的技术，应用在精细胞分离和含有其中所用适体或核酸序列的组合物领域，能够解决费用高精细胞不可逆染色等问题

Inactive Publication Date: 2009-10-07
BIOCERN
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Problems solved by technology

Despite the advantage of being able to control and plan the sex composition of cattle stocks, this method is not more widely used in industry because of the expense of current methods of sorting sperm cells into sex-specific sperm cells using flow cytometry techniques. High and involves irreversible staining of sperm cells prior to insemination

Method used

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  • Sperm cell separation methods and compositions containing aptamers or nucleic acid sequences for use therein
  • Sperm cell separation methods and compositions containing aptamers or nucleic acid sequences for use therein
  • Sperm cell separation methods and compositions containing aptamers or nucleic acid sequences for use therein

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[0062] Sperm aptamer selection method

[0063] The preferred method of selecting specific aptamers of the present invention employs a method that utilizes several rounds of incubation of unsorted, sorted X and / or sorted Y sperm cells with the DNA library and the final step is to collect the solution with the DNA binder and to The solution was used as a template for PCR and strand separation methods on streptavidin beads. Applicants have determined that more than 6 rounds of sperm aptamer selection according to the invention are optimal. Preferably 7, more preferably 8 and even more preferably 9 rounds of selection are performed as described below. However, if necessary, more than 9 rounds of selection can be performed to obtain aptamers that bind to specific target molecules. It is understood that the protocols described below can be varied in temperature, volume and other parameters that do not affect the results of the protocol but result in the production of specific apta...

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Abstract

The present invention provides an aptamer or pool of aptamers that bind(s) to a target molecule on the surface of a mammalian sperm cell and a method for producing the aptamers. The method comprises contacting a collection of different nucleic acid molecules with the target molecule under conditions favorable for binding between at least one of the nucleic acid molecules and the target molecule, to form at least one complex comprising the nucleic acid molecule bound to the target molecule, wherein each of the nucleic acid molecules comprises at least one segment of randomized nucleotide sequences. The complexes are then separated from the unbound nucleic acid molecules and unbound target molecules, and the bound nucleic acid molecule is recovered from the separated complex. The aptamers are used to separate sperm cells based upon sperm cell qualities, such as whether the cells contain X chromosomes or Y chromosomes.

Description

[0001] Related applications [0002] This application claims priority to US Provisional Patent Application Serial No. 60 / 824,069, filed August 30, 2006, which is hereby incorporated by reference in its entirety. technical field [0003] The present invention relates to a method for identifying and isolating cells from a population of cells, especially mammalian sperm cells, by using nucleic acids, especially aptamers, that specifically bind with high affinity and specificity or preference to sperm cells containing an X chromosome or a Y chromosome and composition. Background technique [0004] The ability to select sperm cells with desired characteristics remains an important goal of artificial propagation. An efficient and economical method of identifying and isolating sperm cells for sex selection would be of great economic importance to the livestock industry and especially the beef and dairy industries. In the beef industry, for example, bulls are of higher commercial...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/70C07H21/02C12Q1/68
CPCC12N15/115C12N2310/16C12N2320/10C12N5/0612C12Q1/6811C12Q1/6879A61P43/00C12N5/061
Inventor 戴维·A·奥克森伯格谢尔盖·克雷洛夫迈克尔·穆希夫
Owner BIOCERN
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