Method for producing transgenic animals

摘要

The present invention relates to methods for producing transgenic animals using retroviral constructs engineered to carry a transgene(s) of interest.

主权项

1. A method of producing a transgenic mouse comprising:
transfecting a packaging cell line with:
a first retroviral construct comprising the R and U5 sequences from a 5′ lentiviral LTR, a self-inactivating 3′LTR, a transgene of interest, and a promoter operably linked to the transgene of interest;a second retroviral construct comprising the HIV-1 packaging vector with the env, nef, 5′LTR, 3′LTR, and vpu sequences deleted; andan expression plasmid encoding a pseudotyped envelope glycoprotein; recovering recombinant pseudotyped retrovirus from the packaging cell line; infecting an oocyte or single cell embryo with the recombinant pseudotyped retrovirus, wherein the recombinant pseudotyped retrovirus comprises the R and U5 sequences from a 5′ lentiviral long terminal repeat (LTR) and a self-inactivating lentiviral 3′ LTR; and implanting the oocyte once fertilized or single cell embryo in a pseudopregnant female to produce a transgenic mouse, wherein the pseudotyped retrovirus is integrated into the genome of the mouse embryo, and wherein the transgenic mouse is able to pass the transgene of interest to a progeny of the transgenic mouse such that the progeny expresses the transgene of interest.