Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites.
A satellite DNA-based mammalian artificial chromosome (ACes) was generated and subsequently modified by targeting of a loxP-red fluorescent protein (RFP) expression cassette via homologous recombination into a ribosomal DNA (rDNA)-containing locus. Clones containing correctly targeted ACes were identified by PCR from populations of RFP-expressing cells enriched by FACS sorting and were further characterized by fluorescent in situ hybridization. The targeted ACes maintained its ability to be purified to near homogeneity. Studies are currently underway to further characterize the functionality, carrying capacity, stability and transfectability of this modified ACes.
Stewart
S
S
Chromos Molecular Systems Inc., Burnaby, BC, Canada.
MacDonald
N
N
Perkins
E
E
DeJong
G
G
Perez
C
C
Lindenbaum
M
M
D000818:Animals; D022221:Chromosomes, Artificial, Mammalian; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering; D015316:Genetic Therapy; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein