DNA tetrahedron-based split aptamer probes for reliable imaging of ATP in living cells

Accurate detection and imaging of adenosine triphosphate(ATP)expression levels in living cells is of great value for understanding cell metabolism,physiological activities,and pathologic mechanisms.Here,we developed a DNA tetrahedron-based split aptamer probe(TD probe)for ratiometric fluorescence imaging of ATP in living cells.The TD probe is constructed by hybridizing two split ATP aptamer probes(Apt-a and Apt-b)to a DNA tetrahedron assembled by four DNA oligonucleotides(T1,T2,T3 and T4).In the presence of ATP,the TD probe will alter its structure from the open to closed state,thus bringing the separated donor and acceptor fluorophores into close proximity for high fluorescence resonance energy transfer(FRET)signals.The TD probe exhibits low cytotoxicity,efficient cell internalization and good biological stability.Moreover,based on the FRET“off”to“on”signal output mode,the TD probe can effectively avoid false-positive signals from complex biological matrices,which is significant for long-term reliable imaging in living cells.In addition,by changing the split aptamers attached to DNA tetrahedron,the proposed strategy may be extended for detecting various intracellular targets.Collectively,this strategy provides a valuable sensing platform for biomarkers analysis in living cells,thus having great potential for early clinical diagnosis and therapeutic evaluation.

An Improved Turn-On Aptasensor for Thrombin Detection Using Split Aptamer Fragments and Graphene Oxide

An improved tum-on aptasensor for thrombin detection using split aptamer fragments and graphene oxide （GO） was reported. The thrombin-binding aptamer （Aptl5） was split into two parts for target recognition, an 8-base se- quence labeled with fluorescein （FAM-Apt-A） and a 7-base oligonucleotide sequence （Apt-B）. In the absence of target protein, the fluorescence of FAM-Apt-A/Apt-B was quenched by GO through n-n stacking between GO and single-stranded DNA. However, when thrombin was introduced into the system, a target-induced G-quadruplex forms with two split aptamer fragments and thrombin. The fluorescence recovered due to weak interaction between G-quadruplex and GO. Compared to the strategy using intact aptamer, probe concentration was lowered, and an improved sensitivity was obtained. Moreover, heating process to avoid unfavorable secondary structure was avoided due to the use of shorter split aptamer fragments.

A Sandwich-type Lateral Flow Strip Using a Split, Single Aptamer for Point-of-Care Detection of Cocaine

The sandwich-type lateral fl ow assays relying on dual aptamers with high sensitivity and specifi city have been broadly explored.However,it is unlikely to match a pair of specifi c aptamers that can bind a small molecular target(e.g.,cocaine)simultaneously due to the steric hindrance.In response,we herein introduced the strategy of“one divides into two”into the construction of sandwich-type lateral fl ow strip assay(LFSA).Specifi cally,we split a single cocaine-recognizing aptamer into two segments,either of which was conjugated with gold nanoparticles(AuNPs)or labeled with biotin,serving as signal probe and capture probe,respectively.Upon the presence of the target molecule,a ternary sandwich complex comprised of the two halves of the aptamer and the target formed.Such sandwich-type LFSA exhibited an excellent nonlinear logarithmic response in the range from 10μmol/L to 5 mmol/L with R^(2)=0.9994.The sensitive on-site detection of cocaine in artifi cial biological samples including urine and sweat was achieved within 15 min,with the visual limit of detection as low as 50μmol/L for urine and 200μmol/L for sweat,and the recovery rates of 83.6-107.4%.