Development of a background signal suppression probe for 8-oxoguanine DNA glycosylase detection

Unlocking the Full Potential of Cas12a: Exploring the Effects of Substrate and Reaction Conditions on Trans-Cleavage Activity

“…In ssDNA-activated Cas12a systems, an increase in the concentration of crRNA has almost no effect on the reaction rate (Figure S3b), which means a 1:1 Cas12a/crRNA ratio is suitable for ssDNA-activated systems. Enzymatic reactions may also be significantly influenced by the reaction buffer. − Therefore, we explored eight buffers commonly used in the laboratory. Surprisingly, the results showed that Cas12a had the best trans-cleavage activity in NEBuffer 4 (Figures b and S4), which is proximally five times faster than in the recommended buffer NEBuffer 2.1 provided alongside Cas12a by NEB.…”

Molecular beacons with oxidized bases report on substrate specificity of DNA oxoguanine glycosylases

“…24,25 Fluorescent molecular beacons have been established as excellent tools for characterizing glycosylase activity in a simple and real-time manner. [26][27][28][29] However, to our knowledge, such a strategy has not been established as yet to evaluate glycosylase substrate specicity and screen mutants with altered function due to a lack of synthetic probes containing different glycosylase substrates and the typical use of puried glycosylase enzymes.…”

Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine

“…BER pathway generates a leading role in protecting cells in the process of DNA oxidative damage ( 20 ), which contains the following enzymes: 8-hydroxyguanine DNA glycosidase (OGG1), AP site endonuclease 1 (APE1), DNA polymerase β (POL-β), poly(ADP-ribose) polymerase 1 (PARP1), and DNA ligase (DNA ligase III/XRCC1) ( 21 ). In the BER pathway, OGG1 is an important biomarker, which is mainly responsible for identifying and removing 8-oxo-7,8-dihydro-2′-deoxyguanosine/8-OHdG, so as to maintain the integrity of genome function ( 22 , 23 ). OGG1 is necessary to maintain the proliferation of tumor cells, tightly involving in the occurrence and development of multiple cancers ( 24 , 25 ).…”