6 results about "Embryo transplantation" patented technology

This invention relates to a method for constructing a mouse model of autosomal dominant polycystic kidney disease and its applications, particularly to a method for preparing a non-human animal model carrying a nonsense mutation in the Pkd1 gene c.616_618GAG>TAG based on CRISPR / Cas9 gene editing technology. The method involves co-injecting mouse zygotes with gRNA targeting exon 5 of the Pkd1 gene, homologous recombinant donor oligonucleotides containing the c.616_618GAG>TAG mutation, and Cas9 nuclease. Mutant mice are obtained via embryo transfer. The mutation site is verified by PCR combined with sequencing, and a stable mutant line is established through breeding. This invention also includes the application of this model animal as a research tool for autosomal dominant polycystic kidney disease, particularly in the areas of pathogenesis analysis, drug screening, and therapeutic target validation. This model can stably simulate the typical clinical manifestations and pathological features of human autosomal dominant polycystic kidney disease, providing an important tool for basic research and translational medicine of this disease.

This invention provides a method for preparing goats overexpressing the FOXL2 gene, belonging to the field of genetic engineering and transgenic animal technology. The method includes the following steps: transfecting intersex goat embryonic fibroblasts with a vector plasmid overexpressing the FOXL2 gene to obtain a stable monoclonal cell line overexpressing the FOXL2 gene; using the monoclonal cell line as a nuclear donor for nuclear transfer cloning in a recipient goat to obtain cloned goat embryos overexpressing the FOXL2 gene; and transferring the cloned goat embryos into a surrogate goat to produce goats overexpressing the FOXL2 gene. The nucleotide sequence of the FOXL2 gene is shown in SEQ ID NO.1. By overexpressing the FOXL2 gene in intersex goats, the regulatory function of the FOXL2 gene on horn traits is explored, providing insights for reducing or even overcoming the harm of intersex hornless syndrome and obtaining normal hornless dairy goat germplasm.

The application discloses a SNP molecular marker for identifying the gender of sheep and application thereof. The SNP molecular marker comprises one of SNP1, SNP2, SNP3, SNP4 and SNP5. The application provides a SNP molecular marker for identifying the gender of sheep, and the SNP molecular marker has high typing quality, single copy and high sample detection rate, and can be used for molecular marker assisted breeding of the gender of sheep. The SNP molecular marker can realize early gender typing identification of sheep embryos in a quick, high-throughput and low-cost manner, and the gender identification can be completed before embryo transplantation, so that pregnancy, delivery and feeding consumption of non-target embryos can be effectively avoided, and the breeding cycle can be directly shortened, and the selection efficiency is improved.

The application provides a method for judging endometrial receptivity. The method comprises the following steps: extracting total RNA of endometrium related tissue sample; reverse transcribing the total RNA of the sample into cDNA and amplifying and constructing a library; performing high-throughput sequencing on the library to obtain transcriptome data, analyzing and obtaining the expression level of genes related to endometrial receptivity; inputting the expression level of the related genes into an endometrial receptivity prediction model, and judging the endometrial receptivity according to the calculation result. The method can quickly and accurately judge the endometrial receptivity, and predict the endometrium related tissue of the subject suitable for the implantation window period of embryo transplantation.

The present application relates to a method for constructing a pig endogenous retrovirus inactivated xenotransplant donor pig, and belongs to the technical field of xenogenic organ transplantation. The copy number and PERV-pol genotype of PERVs are determined by ddPCR, Sanger sequencing and second-generation sequencing. Based on the RNP system of CRISPR / Cas9, the pig PERV-pol gene targeting sequence is designed and synthesized, and is co-transfected into a pig fetal fibroblast cell line with spCas9 protein, positive cells are screened, somatic cell cloning and embryo transfer are carried out, and the PERV-pol knockout condition of the fetus is identified when pregnancy occurs. If not completely knocked out, the sgRNA targeting sequence needs to be designed and synthesized again, and the cell line with completely knocked out PERV-pol gene is screened. If completely knocked out, somatic cell cloning and embryo transfer are directly carried out; and the xenotransplant donor pig with inactivated PERVs is obtained.