Molecular beacon probes for the detection of cisplatin-induced DNA damage

Abstract

Cisplatin (cis-diamminedichloroplatinum(II)) causes crosslinking of DNA at AG and GG sites in cellular DNA, inhibiting replication, and making it a useful anti-cancer drug. Several techniques have been used previously to detect nucleic acid damage but most of these tools are labour-intensive, time-consuming, and/or expensive. Here, we describe a sensitive, robust, and quantitative tool for detecting cisplatin-induced DNA damage by using fluorescent molecular beacon probes (MB). Our results show a decrease of fluorescence in the presence of cisplatin-induced DNA damage, confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The decrease in fluorescence upon damage scales with the number of AG and GG sites, indicating the ability of MB to quantitatively detect DNA damage by cisplatin.

An Oligonucleotide‐Distortion‐Responsive Organic Transistor for Platinum‐Drug‐Induced DNA‐Damage Detection

“…The ultralow LOD of our methods is about 5 orders of magnitude lower than that reported for the current DNA‐damage detection methods. [ 4a–e,5,13 ] In addition, a rapid response speed is also essential for high‐throughput damage analysis. Current DNA‐damage sensors mostly require extra process for highly efficient DNA–drug binding and signal amplification, which would bring increased time cost.…”

“…By DROT‐based direct monitoring of the DNA damage process, the signal change can reach a saturation state with ≈20.5 ± 8.7 s after adding drug, which is even shorter than that of similar strategy using time‐lapse AFM imaging. [ 13f ] With steady‐state monitoring time included, a total analysis time of only ≈15 min is adequate for DNA‐damage analysis by nine different conditions.…”

Combining Inquiry-Based and Team-Teaching Models to Design a Research-Driven, Cross-Disciplinary Laboratory Course

“…At this point, every group started by determining what effect cisplatin would have on their beacons by following the exact fluorescence testing protocol from the Shire and Loppnow paper. 1 In nearly all cases, groups observed a significant decrease in signal for the trials where the target sequences had been incubated with cisplatin (Figure 8). With their "baseline" established, groups then compared the signal of the trials where they incubated the target sequences with their analogues.…”

“…Students would then take this new course after completing their General Chemistry, Organic Chemistry, and Inorganic Chemistry sequences and concurrently with their Biochemistry sequence. Finally, the department also used this as an opportunity to incorporate emerging and cutting-edge techniques 1 in this new course, instead of basing it on more traditional "canned" lab exercises.…”

Molecular Beacon Probes of Oligonucleotides Photodamaged by Psoralen

“…Time‐resolved fluorescence has also been used to detect DNA damage (20). Recently, molecular beacons (MBs) have been used as sensitive and accurate probes of UV‐induced, chemical and oxidative DNA damage (21–24; A. Mah, F. Teimoory and G.R. Loppnow, unpublished data; Z. Shire and G.R.…”