Analysis of mammalian telomere position effect.
Methods relating to the positioning of a transgene next to a newly formed telomere in human (HeLa) cells and the subsequent analysis of the resulting clones are described. These include vector design, analysis of integration sites by Southern blotting, pharmacological relief of silencing, and enhancement of silencing by telomere elongation. Several potential pitfalls of applying these techniques to other cell lines are discussed. In addition, detailed instructions are provided for several more general methods related to human telomeres including terminal restriction fragment analysis and purification of telomeres from digested genomic DNA. This chapter summarizes the techniques currently in use that relate to human telomere position effect.
Baur
Joseph A
JA
Department of Cell Biology, The University of Texas Southwestern Medical Center, Dallas, TX, USA.
Wright
Woodring E
WE
Shay
Jerry W
JW
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D020868:Gene Silencing; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008322:Mammals / Q000235:genetics*; D016615:Telomere / Q000235:genetics*; D019076:Transgenes
D004247:DNA