Patent ID: 7101541
Filing Date: 2006-09-05
Classification: A61K,C12N

Abstract:
1. A retroviral vector for gene reconstitution in vitro, comprising: a 3′ portion of a heterologous nucleic acid sequence of a gene to be reconstituted; a 5′ portion of the heterologous nucleic acid sequence of the gene to be reconstituted; and a viral nucleic acid comprising a R, U5 and U3 region, wherein (i) the combined 3′ portion of the heterologous nucleic acid sequence and the 5′ portion of the heterologous sequence have a size of less than seven kilobases, (ii) the 5′ end of the 3′ portion of the heterologous nucleic acid sequence comprises at least a 12 nucleotide direct repeat of the 3′ end of the 5′ portion of the heterologous nucleic acid sequence, (iii) the 3′ portion of the heterologous nucleic acid sequence is linked to the viral nucleic acid at the 5′ end, adjacent to the R and U5 regions, wherein the U5 region comprises a first att site (iv) the 5′ portion of the heterologous nucleic acid sequence is linked to the 3′ end of the viral nucleic acid at the 3′ end, adjacent to the U3 region, wherein the U3 region comprises a second att site, and (v) transformation of a eukaryotic cell with the retroviral vector results in reconstitution and duplication of the heterologous nucleic acid sequence through primer template switching during reverse transcription and subsequent integration in the eukaryotic cell's genome.