Patent ID: 8709754

Claim:
A method for providing stable, long-term expression of a transgene in chromatin, comprising: transfecting a vector into a host cell, said vector comprising: a DNA molecule consisting of an isolated mammalian c-myc insulator element (MINE) of a eukaryotic 5′ constitutive DNA I-hyperacelylated domain located approximately 2.5 kb upstream of the c-myc transcription initiation site of the c-myc gene locus, wherein said isolated MINE is a 1.6-kb fragment from upstream of the c-myc exon 1; wherein said mammalian c-myc MINE comprises, in 5′ to 3′ order, a mammalian c-myc barrier element (BE), a mammalian c-myc silencing element 1 (S1), a mammalian c-myc CCCTC-binding factor (CTCF) element, and a mammalian c-myc silencing element 2 (S2), a promoter domain; and a transgene, wherein the promoter domain and transgene are operably linked, and wherein the DNA molecule flanks the promoter and transgene; and where upon transfection the vector is linearized; wherein, following the transfecting step, the linearized vector is integrated into a host chromosome, wherein c-myc MINE when flanking the gene inserted into said chromosome insulates the transcriptional expression of said gene from one or more cis-acting regulatory sequences in chromatin into which the gene has been inserted, thereby providing stable, long-term expression of the transgene.