Construction Method for Immunodeficient Rat Model

A construction method for an immunodeficient rat model, comprising the following steps:

• (1) obtaining a gRNA targeted for knocking out a Prkdc gene of a rat DNA via in-vitro transcription and mixing the gRNA targeted for knocking out the Prkdc gene of the rat DNA with an in-vitro transcribed CAS9 mRNA, then freezing in RNase-free ultrapure water to obtain an injection A;
• obtaining a gRNA targeted for knocking out an IL2Rγ gene of the rat DNA via in-vitro transcription, and mixing the gRNA targeted for knocking out the IL2Rγ gene of the rat DNA with another in-vitro transcribed CAS9 mRNA, then freezing in RNase-free ultrapure water to obtain an injection B;
• (2) amplifying and purifying a human SIRPα genomic DNA, and freezing in RNase-free ultrapure water to obtain an injection C;
• (3) injecting the injection A and the injection B into cytoplasm of fertilized eggs of different rats by a microinjection method, and then transplanting the fertilized eggs into different pseudo-pregnant female rats, and breeding a second-generation rat A and a second-generation rat B;
• (4) injecting the injection C into a pronucleus of a fertilized egg of a rat by microinjection method, and then transplanting the fertilized egg of the rat into a pseudo-pregnant female rat to obtain a second-generation rat C;
• and
• (5) hybridizing the second-generation rat A, the second-generation rat B and the second-generation rat C to breed an F1 generation;
• further intercrossing the F1 generation to obtain IL2Rγ and Prkdc gene knockout immunodeficient rat models with human SIRPα gene.