Multiplexed single-molecule analysis of human telomerase synthesizing DNA
Hentschel, J.; Choi, J.; Bagshaw, C. R.; Lapointe, C. P.; Wang, J.; Jansson, L. I.; Badstubner, M.; Puglisi, J. D.; Stone, M. D.
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Genomic stability in proliferating cells critically depends on telomere maintenance by telomerase reverse transcriptase. Here we developed a real-time single-molecule RNA sequencing approach that visualizes telomerase catalysis and structural dynamics at single-nucleotide resolution using FRET and zero-mode waveguides. The method permits direct detection of dynamic steps and structural states throughout the telomerase catalytic cycle and can be generalized to other nucleic acid polymerase systems.
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