Mouse in-vitro fertilization breeding sex selection method

Abstract

The invention relates to a mouse in-vitro fertilization breeding sex selection method, which comprises the following steps: inserting a luciferase gene into a Y chromosome of a male mouse by utilizing a gene editing technology, and carrying out embryo injection and transplantation to obtain a Lu-Y male mouse through gene identification; adding a substrate of luciferase into a sperm preserving fluid of a Lucy-Y male mouse, where Lucy-Y sperms emit fluorescence with the wavelength of 560 nm, and normal X sperms do not have fluorescence; sorting lucc-Y sperms and normal X sperms through a flow cytometry sorter; observing the sorted luc-Y sperms and normal X sperms by using a fluorescence microscope; judging the proportion of Luc-Y sperms in normal X sperms through fluorescence; and if the proportion exceeds the standard, putting the normal X sperms into the flow cytometry sorter again for sorting until the purity of the normal X sperms reaches the standard. According to the invention, the flow sorting result can be conveniently and intuitively judged through naked eyes, and the screening precision of the X sperms and the Y sperms of the mice is effectively improved.

Description

Technical field: [0001] The invention relates to the technical field of biological experiments, in particular to a sex selection method for in vitro fertilization and breeding of mice. Background technique: [0002] Mouse in vitro fertilization for purification and multiplication. Generally, natural fertilization will result in half male and half male offspring. But different experiments have selective requirements for the sex of the animals. For example, large-scale expansion requires a larger number of female mice, while some animal models require male mice for experiments. Therefore, in order to obtain the desired number of mice of a specific gender, the natural combination of mouse sperm and egg cells will double the workload and cost of breeding, waste manpower and material resources, and it is also against animal ethics to kill mice of undesired genders. Therefore, we developed a sex-selective method in the process of in vitro fertilization of mice, so as to quickly...

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to provide a sex selection method for in vitro fertilization and breeding of mice, which solves the problem of using a flow cytometer to sort the semen of mice directly to separate X sperm and Y sperm in the prior art. Small size, the sorted sperm are prone to be mixed, there is no simple quality control method for testing the purity of the sperm, and the screening accuracy is limited

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