INVENTION NOVELTY
This invention is a new assay termed ADDIT (Addition of de novo initiated telomeres) that can rapidly identify effectors of telomere length, even if the regulators are essential genes and a method of regulating telomere length with a PARP inhibitor.
VALUE PROPOSITION
The researchers developed an in vivo telomere elongation assay where telomerase repeat addition can be monitored over 48 hours. Several analytical procedures for the quantification of telomerase activity, methods for detecting telomerase activity, as well as for identifying compounds that regulate or affect telomerase activity has been reported. However, these methods are time consuming, labor intensive, PCR dependent and susceptible to inhibition by extracts of clinical samples and are difficult to quantify telomerase activity and are subject to false positives. This invention could provide the following advantages:
•Provide insights into telomere length homeostasis.
•Identify potential targets for future therapeutics.
•Identify genes required for telomerase-dependent telomere elongation by measuring de novo telomere addition at a single chromosome over just one cell cycle.
•Allow dissection of new pathways of telomere length regulation.
•The 48 hours required for ADDIT assay allows for identification of essential genes for roles in telomere length regulation.
TECHNICAL DETAILS
Johns Hopkins researchers have developed a de novo telomere elongation assay in mouse cells and with this assay demonstrated that ATM kinase pathway regulates the elongation of telomeres by telomerase. The ADDIT assay was designed in mouse CASTEiJ cells that have telomere length and distribution very similar to humans. This allows functional probing of telomeres in a setting similar to human telomere length regulation.
