A method for increasing the pregnancy rate of recipients of mammalian cloned embryos

A technology for cloning embryos and pregnancy rate, applied in the biological field, can solve the problems of unseen regulation of the pregnancy rate of mammalian receptors in cloned embryos, and achieve the effect of improving the pregnancy rate

Active Publication Date: 2021-08-31
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] However, there is no research report on the regulation of RTL1 gene in donor cells to improve the pregnancy rate of mammalian recipients of cloned embryos

Method used

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  • A method for increasing the pregnancy rate of recipients of mammalian cloned embryos
  • A method for increasing the pregnancy rate of recipients of mammalian cloned embryos
  • A method for increasing the pregnancy rate of recipients of mammalian cloned embryos

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This example is used to illustrate that RTL1 gene silencing can cause developmental arrest of porcine iPS cell nuclear transfer cloned embryos after implantation.

[0038] In this example, the RTL1 gene (RTL1-iPSCs) was overexpressed in the pig iPS cells of nuclear transfer donors, and the RTL1-iPSCs that were consistent with the expression dose of RTL1 in fibroblasts were selected as donors for nuclear transfer by Q-PCR detection cell. We implanted RTL1-iPSC nuclear transfer cloned embryos into 10 recipient sows, and the results of ultrasound examination on the 25th day of pregnancy showed that 9 of them were pregnant. Among them, we compared the fetuses of two pregnant recipient sows with those of pig iPS cell clones without RTL1 overexpression. The first RTL1-iPSC nuclear transfer clone embryo recipient sow was pregnant with 7 RTL1-iPSC nuclear transfer clone fetuses, of which 4 had normal morphological characteristics and 3 were aborted. Another RTL1-iPSCs nuclear...

Embodiment 2

[0049] This example is used to illustrate whether the normal expression of the RTL1 gene not only affects the development of porcine iPS cell nuclear transfer cloned embryos, but also has a conservative regulatory effect on other types of pig nuclear transfer donor cell cloned embryos and cloned embryos of other species.

[0050] By analyzing a recent report on porcine fibroblast cloned pigs (Ruan et al., 2018), it was found that the expression of the RTL1 gene was at a stable low level ( Figure 4 ). At the same time, we also analyzed the transcriptome data of the recently published bovine allantoic tissue cloned by artificial insemination and nuclear transfer (Biase et al., 2016). Abnormal expression of the RTL1 gene appeared in 3 groups of nuclear transfer cloned bovine embryos. Considering that most of the nuclear transfer cloned bovine embryos were aborted at this time point, the expression analysis of the RTL1 gene may be the key to identify the efficiency of dairy cow c...

Embodiment 3

[0055] This example is used to illustrate that the RTL1 gene plays an important role in maintaining the developmental potential of mammalian pluripotent stem cells.

[0056]Although 12 years have passed since iPSC technology was first released in 2006, real iPS cells from large animals such as pigs, sheep, and cattle have not yet been obtained (Huang et al., 2011; Li et al., 2011; Wu et al., 2009). So far, only mouse iPS cells have been proved to have the ability of reproductive mosaicism and tetraploid compensation. Previous studies have pointed out that gene expression in the DLK1-DIO3 region is related to the pluripotency of mouse stem cells (Liu et al., 2010; Stadtfeld et al., 2010), but the specific genes or mechanisms have not yet been determined. We analyzed the relevant data on stem cell research published before (Chang et al., 2014; Guo et al., 2016; Li et al., 2017), the expression of RTL1 can be used as a marker for evaluating the developmental potential of mammali...

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Abstract

The invention belongs to the field of biotechnology, and specifically discloses a method for improving the pregnancy rate of mammalian embryo clone recipients. In the present invention, induced pluripotent stem cells (iPS cells) are used as donor cells, and the pregnancy rate of cloned embryos is significantly improved after performing dosage compensation expression of RTL1.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving the pregnancy rate of mammalian cloned embryo recipients. Background technique [0002] Since the birth of the first mammal cloned sheep "Dolly", nuclear transfer (NT) cloning technology has been successfully applied to more than 20 kinds of mammals, but the efficiency of mammalian cloning (defined as birth / reconstituted oocyte survival rate) is still very low, for example, the efficiency of cloning mice is 1-2% (Ogura et al., 2013), the efficiency of cloning monkeys is 1.5% (Liu et al., 2018), the efficiency of cloning pigs is The efficiency is 0.3%, that of cloned sheep is 0.3%, that of cloned horses is 0.8%, that of cloned cows is 1.7%, etc. (Keefer et al., 2015). In other words, about 99% of somatic cell nuclear transfer cloned embryos did not develop into mature individuals, and mass abortions occurred at various stages of pregnancy (Keefer, 2...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87C12N5/10C12N5/16C12N15/00
CPCA01K2207/12A01K2227/108C12N5/0604C12N15/02C12N15/8778C12N2510/00
Inventor 吴森赵要风杜旭光余大为王靖邹惠影冯涛
Owner CHINA AGRI UNIV
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