Efficient signaling platforms built from a small catalytic DNA and doubly labeled fluorogenic substrates

摘要: RNA-cleaving deoxyribozyme 8-17 has been increasingly used in nanotechnology and biosensing applications. Conventional methods to equip with fluorescent signaling property usually involve covalent attachment of two dyes at nucleotide positions that are far away from the catalytic core, such bulky dye structures would not affect activity. However, maximum enhancement associated these constructs is typically ≤10-fold, due a high background. To find an optimal balance between signal speed, we have conducted comprehensive study on effects nature (Alexa Fluor 488, 546 647; QSY 9 21) as well their attaching along substrate strand performance 8-17. Our results indicated able cleave almost every modified substrate, including those chromophores only 1 nt cleavage site. Most importantly, all substrates generate 15- 85-fold within 10 min. We also provided guidelines for selecting could offer best enhancement, fastest or speed.