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Method for screening donor cell line based on blastocyst gene expression level

A gene expression level, donor cell technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of long pregnancy cycle, time-consuming and laborious, etc., and achieve the effect of improving cloning efficiency

Active Publication Date: 2015-04-22
NORTHWEST A & F UNIV +1
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  • Summary
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  • Description
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Problems solved by technology

However, due to the long pregnancy cycle of large animals, it is time-consuming and laborious to screen donor cell lines that are easy to be reprogrammed and have high cloning efficiency after the cloned animals are born.

Method used

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  • Method for screening donor cell line based on blastocyst gene expression level
  • Method for screening donor cell line based on blastocyst gene expression level

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Embodiment Construction

[0031] The present invention provides a method for screening donor cell lines based on gene expression levels on blastocysts. After nuclear transfer of donor cell lines from different individuals, TCF7, DGCR8, SENP7, DNMT3B, Vimentin, IFI6, The expression levels of EFNA2, F5, IRAK1, PRICKLE1, TNFRSF1A, ARG2, NEK6, OCT4, NANOG, SOX2, CDX2, XIST and IGF2R were selected on cloned blastocysts. The expression levels of these genes were all the same as those on normally developed in vitro fertilized blastocysts. The donor cell line whose expression level is not significantly different is a nuclear donor cell, which can significantly improve the cloning efficiency of bovine somatic cells.

[0032] In the following, the present invention will be further described in detail with reference to the specific operation process and comparative analysis results, which is an explanation rather than a limitation of the present invention.

[0033] The methods and operations for screening donor cell l...

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Abstract

The invention discloses a method for screening a donor cell line based on blastocyst gene expression level. The method comprises the steps of: detecting expression levels of TCF7, DGCR8, SENP7, DNMT3B, Vimentin, IFI6, EFNA2, F5, IRAK1, PRICKLE1, TNFRSF1A, ARG2, NEK6, OCT4, NANOG, SOX2, CDX2, XIST and IGF2R at a blastula stage after carrying out nuclear transferring on the donor cell lines from different individuals; and selecting the donor cell line which is not significant in difference of all expression levels of these genes on cloned blastocysts and the expression level of normally growing in vitro fertilization blastocyst as a nuclear donor cell. The cloning efficiency of bovine somatic cells can be significantly improved.

Description

Technical field [0001] The invention belongs to the technical field of animal somatic cell cloning, and relates to a method for screening donor cell lines based on the expression level of blastocyst genes. Background technique [0002] Somatic cell cloning is a technology with great research and commercial value. It can be applied to therapeutic cloning, disease models, human organ transplantation, animal transgenic research, protection of endangered animal breeds, and expansion of excellent livestock breeds. However, the efficiency of somatic cell cloning is still not high, which significantly restricts the wide application of this technology. Among them, the genetic background of the donor cell has a great influence on the cloning efficiency of animals. Some cell lines are used for nuclear transfer, and the final cloning efficiency is very high, and the born cloned animals do not show obvious developmental abnormalities; but some cell lines do The vitality is very good, and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/873C12Q1/68
Inventor 张涌苏建民王勇胜刘军
Owner NORTHWEST A & F UNIV
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