Human repeat-mediated integration of selectable markers into somatic cell hybrids.
We describe a strategy to introduce preferentially the dominant selectable marker neoR into the human chromosome within a monochromosome hybrid cell line. Integration of a construct containing the marker is mediated by human-specific repeat elements that promote multilocus human-specific integration with a single targeting vector. We tested two classes of repeat elements: the Alu family of SINE repeats and the Line1 repeat family. We show that Alu sequences alone are insufficient to direct human-specific integration but when used in combination with a Line1 element, or when only Line1 elements are included, integration of the vector into the human component of a monochromosome somatic cell hybrid is favored. The vectors also carry sequences that facilitate mapping and selective cloning of the targeted region. This strategy provides a means to generate selectable human subchromosomal fragments that can be used for localization of genes through positional cloning and, more important, for the identification of functional units through DNA transfer.
Watson
J E
JE
MRC Human Genetics Unit, Western General Hospital, Edinburgh, UK.
Slorach
E M
EM
Maule
J
J
Lawson
D
D
Porteous
D J
DJ
Brookes
A J
AJ
D000900:Anti-Bacterial Agents; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D018390:Gene Targeting; D005819:Genetic Markers; D005822:Genetic Vectors; D005839:Gentamicins; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection
D000900:Anti-Bacterial Agents; D005819:Genetic Markers; D005839:Gentamicins; D004247:DNA; C010680:antibiotic G 418