Vectors for Gene Transfer Derived from Animal DNA Viruses: Transient and Stable Expression of Transferred Genes

Establishment of an oriP/EBNA1-based episomal vector transcribing human genomic β-globin in cultured murine fibroblasts

“…The blot was probed with an AlfIII-digested pBH140 vector that was used to generate the 200-kb pBH148. 10 All the A9pBH148 episomes iso- lated from any culture to week 12 (lanes [5][6][7][8][9][10][11][12][13][14] migrated at approximately the same position as the episome from the positive control DRpBH148 (lane 1). A DNA ladder, from the ethidium bromide-stained gel, is superimposed on the Southern blot to illustrate the distances between the intact 200-kb pBH148 circular episome, any sheared linear pBH148, and genomic fragments.…”

“…[2][3][4][5] Self-replicating viral-based vectors are capable of longterm episomal persistence in mammalian cells. 6,7 These vectors contain components derived from (1) EpsteinBarr virus (EBV; oriP/EBNA1 viral elements); [8][9][10] (2) bovine papilloma virus (BPV-1; ori elements); 11,12 (3) genomic components from either yeast (YACs) or human chromosomes (HACs); [13][14][15][16] and (4) chimeric viral/genomic systems either engineered from the oriP/EBNA1-based pLIB or from mammalian artificial episomal chromosomes (MAECs). [17][18][19] The advantages and limitations of these various viral vector-based systems in biological research are discussed elsewhere.…”