Cancers must utilize a telomere maintenance mechanism (TMM) to overcome telomere shortening and achieve replicative immortality. While 85% of cancers express telomerase to lengthen their telomeres via reverse transcription, the remaining 15% utilize a homologous recombination-based pathway termed the alternative lengthening of telomeres (ALT). The biological mechanics of ALT are not fully characterized, and ALT cancers are typically severe with a poor prognosis, so there is a clinical need to further understand and target the ALT pathway. The purpose of this research was to develop a high throughput assay for detecting C-circles, a quantifiable phenotypic marker unique to ALT cancers and tightly associated with ALT activity, based off a single tube assay in literature. The assay was optimized to lower costs and decrease total time while maintaining a significant signal over background by modifying incubation times for cell lysis, DNA amplification steps, and DNA polymerase enzyme concentration for robust amplification. The high throughput assay has been validated and will be implemented for compound screening.
C. L. Cook (Sat,) studied this question.
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