The object of the present invention is to provide a test method using a novel administration method of an extract from inflamed tissues inoculated with vaccinia virus or a preparation containing the extract. It has been demonstrated that by sustainedly administering an extract from inflamed tissues inoculated with vaccinia virus or a preparation containing the extract by using a continuous administration device according to the present invention, a comparable effect is exerted at a very low dose compared with conventional administration methods such as oral administration. Therefore, the present invention can provide a test method using animals requiring less burden on a researcher, a treatment of patient at a low dose, and the like.

Method for selecting mammalian cells having a genetically-desirable trait, the method comprising culturing an embryo in vitro, dividing the cells from the embryo into aliquots, subjecting the cells from at least one of the aliquots to a genetic analysis, and, based on the results of such analysis, selecting an aliquot of cells. Method for selecting mammalian cells having a genetically-desirable trait, the method comprising culturing an embryo in vitro, transferring the cultured embryo into a recipient female, collecting the embryo, dividing the cells from the embryo into aliquots, subjecting the cells from at least one of the aliquots to a genetic analysis, and, based on the results of such analysis, selecting an aliquot of cells. Method for selecting a mammalian embryo having a genetically-desirable trait, the method comprising removing one or more cells from each of a plurality of embryos, culturing the cells, subjecting the cultured cells to a genetic analysis, and, based on the results of such analysis, selecting an embryo.

A method of modulating some or all copies of a gene in a cell is provided including introducing into a cell one or more ribonucleic acid (RNA) sequences that comprise a portion that is complementary to all or a portion of each of the one or more target nucleic acid sequences, and a nucleic acid sequence that encodes a Cas protein and maintaining the cells under conditions in which the Cas protein is expressed and the Cas protein binds and modulates the one or more target nucleic acid sequences in the cell.

Genetic tests, such as whole genome analysis (WGA), have been employed to identify genetically superior embryos. The disclosed methods extend in vitro culture time of embryos while awaiting results of genetic tests being performed on a portion of the same embryos. The disclosed methods also help expand the number of cells in each embryo before implantation in the recipient.

Disclosed herein, are inducible immunodeficient animals and methods to make them by adding an IL2Rg/RAG2 rescue cassette (RG-reg) or an IL2Rg/RAG2/FAH rescue cassette (FRG-reg) to a line of IL2Rg/RAG2 knockout (RG-KO) or IL2Rg/RAG2/FAH knockout (FRG-KO) swine. The rescue cassette enables line breeding of immunocompetent (regRG-KO) or (regFRG-KO) swine for rapid propagation. The rescue cassette can be excised, specifically in germ cells of regRG-KO or regFRG-KO swine, such that offspring of animals do not possess the rescue cassette and are immunodeficient. The immunodeficient swine also provide host embryos having genetic ablations to provide a niche for organ complementation by human stem cells.

Provided for the first time in the present invention is a method for preparing a non-human primate somatic cell cloned animal, which method specifically comprises the steps of: (i) providing a reconstructed egg, wherein the egg comes from the non-human primate (ii) activating the reconstructed egg to form an activated reconstructed egg or activated reconstructed embryo formed by the reconstructed egg; (iii) reprogramming (a) the activated reconstructed egg or (b) embryonic cells of the activated reconstructed embryo to obtain a reprogrammed reconstructed egg or reprogrammed reconstructed embryo; and (iv) regenerating the reprogrammed reconstructed egg or reprogrammed reconstructed embryo to obtain the non-human primate somatic cell cloned animal. The method of the present invention can significantly improve the developmental capacity of nucleus-transplanted embryos in non-human primates (such as monkeys).

The present invention relates, inter alia, to methods for determining whether or not an animal and/or its offspring is more likely than not to have an increased tolerance to heat, an increased resistance to ticks, and/or a desirable coat texture. It also provides methods for selecting or rejecting animals, one or more cells or embryos, estimating the worth of an animal, generating animals having a desired genotype/phenotype, cloning and breeding animals and herd formation.

Provided is a double-gene knockout vector system, a method for preparing porcine fibroblasts with both CD163 gene and CD13 gene being knocked-out, prepared porcine fibroblasts, and a method for preparing a gene-edited pig with both CD163 gene and CD13 gene being knocked-out. The vector system of the present disclosure comprises a CD163 gene knockout vector and a CD13 gene knockout vector. The CD163 gene knockout vector comprises a gene editing vector backbone and a DNA fragment ligated to the gene editing vector backbone, with a nucleotide sequence of the DNA fragment being shown in any one of SEQ ID NOs: 1-3. The CD13 gene knockout vector comprises a gene editing vector backbone and a DNA fragment ligated to the gene editing vector backbone, a nucleotide sequence of the DNA fragment being shown in any one of SEQ ID NOs: 4-6.

The present invention relates, inter alia, to methods for determining whether or not an animal and/or its offspring is more likely than not to have an increased tolerance to heat, an increased resistance to ticks, and/or a desirable coat texture. It also provides methods for selecting or rejecting animals, one or more cells or embryos, estimating the worth of an animal, generating animals having a desired genotype/phenotype, cloning and breeding animals and herd formation.

The invention encompasses methods for increasing genetic progress in livestock, and for genetic dissemination, including the use of amniocentesis to obtain fetal amniocytes for use in genomic evaluation and cloning.