IRES mediated multicistronic vectors

摘要

This invention relates to nucleic acid molecules comprising at least one nucleic acid sequence encoding for a peptide or protein of interest, at least one nucleic acid sequence encoding for a selectable marker, and at least one IRES sequence, wherein the at least one IRES sequence is located between the at least one nucleic acid sequence encoding for the peptide or protein of interest and the at least one nucleic acid sequence encoding for the selectable marker. Furthermore, this invention relates to host cells comprising such nucleic acid molecule and to methods of recombinant protein expression using such host cells.

主权项

1. An isolated nucleic acid molecule comprising:
at least one nucleic acid sequence encoding for a peptide or protein of interest, at least one nucleic acid sequence encoding for a selectable marker, and at least one internal ribosome entry site (IRES) sequence, wherein the at least one IRES sequence is located between the at least one nucleic acid sequence encoding for the peptide or protein of interest and the at least one nucleic acid sequence encoding for the selectable marker, and the at least one nucleic acid sequence encoding for a selectable marker has a reduced translation efficiency and/or the encoded selectable marker is mutated such that it has a lowered activity compared to its wildtype variant, wherein the at least one IRES sequence comprises a nucleic acid sequence selected from the group of the nucleic acid sequences set forth in SEQ ID NO:02-57 and 83-112, wherein the at least one IRES sequence has a mutation that reduces ribosome binding affinity compared to the corresponding wildtype IRES sequence, and wherein the encoded selectable marker is attenuated.