Analysis of Average Telomere Length in Human Telomeric Protein Knockout Cells Generated by CRISPR/Cas9.
Telomeres play an important role in ensuring the integrity of the genome. Telomere shortening can lead to loss of genetic information and trigger DNA damage responses. Cultured mammalian cells have served as critical model systems for studying the function of telomere binding proteins and telomerase. Tremendous heterogeneity can be observed both between species and within a single cell population. Recent advances in genome editing (such as the development of the CRISPR/Cas9 platform) have further enabled researchers to carry out loss-of-function analysis of how disrupting key players in telomere maintenance affects telomere length regulation. Here we describe the steps to be carried out in order to analyze the average length of telomeres in CRISPR-engineered human knockout (KO) cells (TRF analysis).
Cell-based Assay Screening Service Core, Baylor College of Medicine, Houston, TX, 77030, USA.
Jun
J
Xu
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA.
Zhou
Z
Songyang
Cell-based Assay Screening Service Core, Baylor College of Medicine, Houston, TX, 77030, USA.|Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA.
Dan
D
Liu
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA. hyeungk@bcm.edu.
Hyeung
H
Kim
D064113:CRISPR-Cas Systems / Q000502:physiology*; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000502:physiology; D006801:Humans; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000378:metabolism*; D034501:Telomere-Binding Proteins / Q000378:metabolism
D034501:Telomere-Binding Proteins; D019098:Telomerase
CRISPR; Cas9; Knockout; TRF; Telomere length; Telomere maintenance