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Construction of a POT1 conditional knock out in human HCT116 cells
Author(s) -
Junaid Yetunde,
Rajkisore Shivika,
Loayza Diego
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1036-c
POT1 is the overhang binding protein at telomeres. POT1 is conserved in evolution and is critical in regulating both telomerase‐mediated telomere maintenance and telomere protection. This study describes the construction of a conditional human somatic cell knock out of POT1, in order to carry out a detailed genetic analysis of POT1 function at telomeres. The experimental setup aimed at studying the short term and long term effects of POT1 loss, and at examining the rescue by specific alleles or domains of POT1 or other candidate proteins in the pathway. The regulation of telomere length was analyzed over time and the status of telomere protection assessed by the measure of the amounts of telomere‐dysfunction induced foci (TIFs) upon POT1 depletion. The results will be put in the context of the contribution of telomere function to chromosome integrity and cell growth.