ALPHAVIRAL VECTORS AND CELL LINES FOR PRODUCING RECOMBINANT PROTEINS

摘要

The present invention relates to polynucleotides and alphaviral vectors for the expression of genes of interest in mammalian cells. Additionally, the invention relates to cells which comprise said polynucleotides and alphaviral vectors and are capable of stably expressing one or more genes of interest. The invention also relates to methods for obtaining said cells, to methods for expressing a gene of interest in said cells, and to methods for replacing the gene of interest stably expressed by said cells with another gene of interest.

主权项

1. A polynucleotide comprising
(i) a transcription regulatory sequence, (ii) a DNA sequence complementary to an alphavirus replicon which is operatively bound to said transcription regulatory sequence, and wherein said alphavirus replicon complementary sequence comprises a first recognition sequence for a site-specific recombinase located between the alphavirus replicon subgenomic promoter and the alphavirus replicon 3′ untranslated sequence, (iii) a transcription termination sequence in 3′ position with respect to the 3′ end of the sequence complementary to an alphavirus replicon, and (iv) a second recognition sequence for a site-specific recombinase located in 3′ position with respect to the transcription termination sequence.