Novel method for introducing slow virus vector into chick embryo

Abstract

The invention relates to a novel method for introducing a slow virus vector into a chick embryo, which comprises the following steps: (1) taking out an early embryo egg (13-17HH) hatched for 48-58h, lightly knocking an eggshell blunt end opening under sterile conditions by using tweezers, and lightly peeling off a shell membrane covered on the upper layer of the embryo by using ophthalmic tweezersafter finding the embryo position, exposing the embryo; (2) diluting the introduced slow virus vector to a fixed titer by using a cell culture medium or a cell buffer solution, and gently dropwise adding the diluted vector aiming at the exposed embryo, so that the embryo is covered with a slow virus solution; and (3) adding 200ul of mycillin to an injection part after the dropwise addition is finished, and then sealing by using a medical adhesive tape to finish the introduction of the slow virus carrier into the chick embryo, and after the sealing is tight, continuously putting into an incubator, and taking out for detection at different stages according to the requirements of the next step. The method can be used for production research of chicken embryo exogenous gene introduction, exogenous gene knockdown or knockout, transgenic chicken and the like.

Description

technical field [0001] The invention relates to a new method for introducing lentiviral vectors into chicken embryos, and belongs to the field of biotechnology. Background technique [0002] As a classic model of embryonic biology, the chicken embryo has been used to explore embryonic development and embryonic cell differentiation, especially in medical models with broad application prospects. The introduction of exogenous vectors into chicken embryos is of great significance to production research such as gene function verification and transgenic breeding. Therefore, an efficient and simple exogenous vector introduction method can not only provide technical support for chicken embryo development biology and transgenic chicken production, but also provide an experimental basis for related research. At present, there have been a large number of reports on the methods of introducing exogenous vectors into chicken embryos at home and abroad. At present, the main methods are d...

AI Technical Summary

Problems solved by technology

Lentiviral vectors can effectively infect early chicken embryos with their unique infection methods, but there is no systematic optimization and uniform description of the method of introducing lentiviral vectors into chicken embryos, which limits the use of lentiviral vectors in chicken embryos. Further application in developmental research requires further optimization and improvement

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