Journal
CELL DISCOVERY
Volume 3, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/celldisc.2017.34
Keywords
CRISPR/Cas9; inducible knockout; metabolism; POT1 isoform; telomere; telosome/shelterin
Categories
Funding
- National Key Research and Development Program of China [2017YFA0102800, 2017YFA0102801]
- National Natural Science Foundation of China [NSFC 91640119, 81330055]
- Science and Technology Planning Project of Guangdong Province [2015B020228002]
- Guangzhou Science and Technology Project [201605030012]
- NIGMS [GM095599]
- NCI [CA211653]
- CPRIT [RP160462, RP120092]
- Welch Foundation [Q-1673, HL131744]
- C-BASS Shared Resource at the Dan L Duncan Cancer Center (DLDCC) of Baylor College of Medicine [P30CA125123]
- Proteomic and Metabolomic Core Facility [NCI/2P30CA125123-09]
- Dan L Duncan Cancer Center (DLDCC)
Ask authors/readers for more resources
CRISPR/Cas9 technology enables efficient loss-of-function analysis of human genes using somatic cells. Studies of essential genes, however, require conditional knockout (KO) cells. Here, we describe the generation of inducible CRISPR KO human cell lines for the subunits of the telosome/shelterin complex, TRF1, TRF2, RAP1, TIN2, TPP1 and POT1, which directly interact with telomeres or can bind to telomeres through association with other subunits. Homozygous inactivation of several subunits is lethal in mice, and most loss-of-function studies of human telomere regulators have relied on RNA interference-mediated gene knockdown, which suffers its own limitations. Our inducible CRISPR approach has allowed us to more expediently obtain large numbers of KO cells in which essential telomere regulators have been inactivated for biochemical and molecular studies. Our systematic analysis revealed functional differences between human and mouse telomeric proteins in DNA damage responses, telomere length and metabolic control, providing new insights into how human telomeres are maintained.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available