Stability of a functional murine satellite DNA-based artificial chromosome across mammalian species.
A 60-Mb murine chromosome consisting of murine pericentric satellite DNA and two bands of integrated marker and reporter genes has been generated de novo in a rodent/human hybrid cell line (mM2C1). This prototype mammalian artificial chromosome platform carries a normal centromere, and the expression of its beta-galactosidase reporter gene has remained stable under selection for over 25 months. The novel chromosome was transferred by a modified microcell fusion method to mouse [L-M(TK-)], bovine (P46) and human (EJ30) cell lines. In all cases, the chromosome remained structurally and functionally intact under selection for periods exceeding 3 months from the time of transfer into the new host. In addition, the chromosome was retained in three first-generation tumours when L-M(TK-) cells containing the chromosome were xenografted in severe combined immunodeficiency mice. These data support that a murine satellite DNA-based artificial chromosome can be used as a functional mammalian artificial chromosome and can be maintained in vivo and in cells of heterologous species in vitro.
Telenius
H
H
Chromos Molecular Systems Inc., Vancouver, BC, Canada.
Szeles
A
A
Keresö
J
J
Csonka
E
E
Praznovszky
T
T
Imreh
S
S
Maxwell
A
A
Perez
C F
CF
Drayer
J I
JI
Hadlaczky
G
G
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002871:Chromosome Banding; D002875:Chromosomes; D004276:DNA, Satellite / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D008677:Metaphase; D051379:Mice; D016513:Mice, SCID; D008967:Molecular Biology / Q000379:methods*; D009374:Neoplasms, Experimental; D016615:Telomere / Q000235:genetics; D013997:Time Factors; D014407:Tumor Cells, Cultured
D004276:DNA, Satellite