Complementation of the beige mutation in cultured cells by episomally replicating murine yeast artificial chromosomes.

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

“…40 This and two previous studies demonstrated that large virus-based circular therapeutic episomes might soon become a reality. 8,19 In addition, initial research using oriP/EBNA1-based 'mini-EBV' vectors led to the generation of an enhanced EBNA1 variant capable of improved episomal maintenance and transgene expression by a new improved second generation mini-EBV-based vector.…”

“…YACs, with up to 1500-kb of human DNA, replicated as episomes in mouse cells following yeast spheroplast fusion, but were unstable in the absence of selective pressure. 13,14,40,41 Although these YACs contained large human DNA fragments (complete with replication origins) they were devoid of any segregation apparatus. In other experiments, use of microcell fusion demonstrated that as little as 100-kb of human DNA could be maintained episomally in EBNA1-expressing mouse cells in low copy number, presumably by EBNA1 protein binding to the oriP on these vectors.…”

The Beige/Chediak-Higashi Syndrome Gene Encodes a Widely Expressed Cytosolic Protein

“…Consistency of the alignment process was confirmed on a daily basis in two ways: For light scatter, a panel of four polystyrene beads ranging in size from 0.48 to 1.43 m (Polysciences, Inc.) was analyzed, and peak channels were compared with previous results. For log FSC the largest bead appeared just above the 10 3 division on a four log scale (linear channels 1200 -1300) and the smallest beads at or below the 10 1 division (linear channels [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20]. The log SSC peaks ranged from the 10 2 (linear channels 70 -130) down to 10 1 (linear channels [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20].…”

“…We cloned the beige gene using a YAC complementation strategy in which YACs were introduced into bg J /bg J mouse fibroblasts (7). During the course of these studies we isolated two complemented cell lines (195-3 and 195-4), in which the beige gene was overexpressed as determined by Northern analysis (4).…”

Identification of a YAC from 16q24 carrying a senescence gene for breast cancer cells

“…YACs have been transferred e ciently into mouse (Pavan et al, 1990;Pachnis et al, 1990;Huxley et al, 1991;Eliceiri et al, 1991;Strauss and Jaenisch, 1992;Peterson et al, 1993;Perou et al, 1996;Koreth et al, 1999) and Chinese hamster cells (Gnirke et al, 1991) by spheroplast fusion but only a few studies have reported successful transfer into human cells (Wada et al, 1994;Julicher et al, 1997;Murakami et al, 1998). Thus, not every cell line is amenable to this method.…”