A highly sensitive electrochemical aptasensor for cocaine detection based on CRISPR-Cas12a and terminal deoxynucleotidyl transferase as signal amplifiers.

Cocaine is one of the mainly used illegal drugs in the world. Using the signal amplification elements of terminal deoxynucleotidyl transferase (TdT) and CRISPR-Cas12a, a highly sensitive and simple electrochemical aptasensor was introduced for cocaine quantification. When, no cocaine existed in the sample, the 3'-end of complementary strand of aptamer (CS) was extended by TdT, leading to the activation of CRISPR-Cas12a and remaining of very short oligonucleotides on the working electrode. So, the current signal was remarkably promoted. With the presence of cocaine, CS left the electrode surface. Thus, nothing changed following the incubation of TdT and CRISPR-Cas12a and the Aptamer/Cocaine complex presented on the electrode. Consequently, the [Fe(CN)6]3-/4- could not freely reach the electrode surface and the signal response was weak. Under optimal situations, the biosensor revealed a wide linear relation from 40 pM to 150 nM with detection limit of 15 pM for cocaine. The sensitivity of the analytical system was comparable and even better than other reported methods for cocaine detection. The designed method displayed excellent cocaine selectivity. The aptasensor could work well for cocaine assay in serum samples. So, the aptasensor is expected to be an efficient analytical method with broad applications in the determination of diverse analytes.

A bivalent binding aptamer-cDNA on MoS2 nanosheets based fluorescent aptasensor for detection of aflatoxin M1.

To ensure the safety of dairy products, especially milk, and consequently protect human health, accurate and simple analytical techniques are highly necessary to determine the low concentration of aflatoxin M1 (AFM1) as an important carcinogen. Herein, a novel, accurate and simple fluorescent aptasensor was designed for selective detection of AFM1 based on bivalent binding aptamer-cDNA (BBA-cDNA) structure. Moreover, MoS2 nanosheets (MoS2 NSs) were used as the fluorescent quencher and FAM-labeled complementary strand of aptamer (FAM-CS) was applied as a fluorescent probe. In this study, we achieved a new result. Unlike previous studies, in this work, the BBA-cDNA structure was not disassembled in the presence of the target. Therefore, as the AFM1 concentration increased, more targets were attached to the BBA-cDNA structure and as a result, the BBA-cDNA structure/AFM1 could not be placed on the surface of MoS2 NSs, leading to the more fluorescent intensity detection. Under optimized conditions, the developed fluorescent analytical method revealed great selectivity toward AFM1 with a limit of detection (LOD) of 0.5 nM and a linear range from 0.7 to 10 nM. This fabricated aptasensor indicated excellent analytical performance for AFM1 detection in milk samples with LOD of 0.1 nM. Overall, the proposed approach could provide an effective basis for small molecule analysis to guarantee food and human safety using appropriate aptamer sequences.

An optical aptasensor for aflatoxin M1 detection based on target-induced protection of gold nanoparticles against salt-induced aggregation and silica nanoparticles.

Not only intoxications of aflatoxins are significant risk for human beings, but also; the contamination with these toxins affect the economy. Therefore, developing a rapid, precise and inexpensive determination method is vitally important. Here, a colorimetric aptasensor is introduced for the detection of aflatoxin M1 (AFM1) based on the preservation of gold nanoparticles (AuNPs) against NaCl-induced aggregation by detaching of complementary strand of aptamer (CS) from the aptamer-modified streptavidin coated silica nanoparticles (SNPs) following the addition of target. So, the color of sample remains red. While, in the lack of AFM1, salt-induced aggregation of AuNPs occurs and the color of sample becomes purple as the aptamer/CS (dsDNA)-modified SNPs is stable and CS cannot bind to AuNPs. The proposed aptasensor could detect AFM1 in a linear dynamic range, 300-75,000 ng/L, with a detection limit of 30 ng/L. Also, the sensing method was effectively applied for AFM1 recognition in milk samples.

Co-delivery of doxorubicin and aptamer against Forkhead box M1 using chitosan-gold nanoparticles coated with nucleolin aptamer for synergistic treatment of cancer cells.

Herein, a nanotherapeutic delivery method was presented for co-delivery of doxorubicin (DOX) and aptamer against Forkhead box M1 (FOXM1 Apt) to cancer cells. Firstly, the vehicle composed of chitosan (CS)-Gold nanoparticles (AuNPs) conjugate was prepared. Nucleolin aptamer (AS1411) and FOXM1 Apt were loaded onto the CS-AuNPs and formed Aptamers (Apts)-CS-AuNPs. Subsequently, DOX was added to the Apts-CS-AuNPs to obtain the DOX-Apts-CS-AuNPs complex for synergistic treatment of tumor. The data of flow cytometry analysis and fluorescence imaging displayed that the complex was effectively internalized into target cells (A549 and 4T1 cells, nucleolin+) but not into CHO cells as nontarget cells. The results of the MTT assay showed that the complex significantly increased cell mortality in 4T1 and A549 cells compared to CHO cells treated with the complex. The in vivo studies demonstrated that the DOX-Apts-CS-AuNPs complex exhibited more tumor inhibitory effect and less distribution in other organs compared to free DOX.

A smart ATP-responsive chemotherapy drug-free delivery system using a DNA nanostructure for synergistic treatment of breast cancer in vitro and in vivo.

This study demonstrated a chemotherapy drug-free delivery system for breast cancer treatment based on a simple DNA nanostructure composed of sequence 1 containing ATP and AS1411 aptamers and sequence 2 containing antimiR-21. The DNA nanostructure was used for co-delivery of KLA peptide and antimiR-21 as antiapoptotic agents. These therapeutic agents could not be internalised into eukaryotic cells freely which is one of the great features of this targeting platform. The presented delivery system was ATP-responsive, leading to disassembly of the DNA nanostructure in high ATP concentration of cancer cells and restoration of the function of antimiR-21 in these cells. The DNA nanostructure was associated with high cellular uptake by MCF-7 and 4T1 cells due to expression of nucleolin as target of AS1411 on their plasma membranes, while the developed targeting platform could not be internalised into CHO cells because of lack of the active targeting moiety on their surfaces. Furthermore, the results showed that co-delivery of antimiR-21 and KLA peptide using the DNA nanostructure could efficiently prohibit tumour growth in vitro and in vivo and induce a synergistic anticancer activity. Thus, this work provides a new ATP-responsive nanotargeting delivery system and synergistic chemotherapy drug-free regimen for cancer treatment.

An ultrasensitive electrochemical sensing method for detection of microcystin-LR based on infinity-shaped DNA structure using double aptamer and terminal deoxynucleotidyl transferase.

This study develops a novel electrochemical sensing platform for microcystin-LR (MC-LR) detection. This aptasensor comprises the hybridization of double aptamer to its complementary strand (CS) on the surface of electrode and generation of an Infinity-shaped DNA structure in the absence of target by terminal deoxynucleotidyl transferase (TdT). The formation of Infinity-shaped construction leads to the development of an ultrasensitive aptasensor for MC-LR detection. In the presence of MC-LR, double aptamer is dissociated from its CS because of its high affinity for MC-LR and leaves the surface of electrode. Subsequently, no Infinity-shaped structure is formed following the introduction of TdT and a strong current signal is observed. The proposed method was employed for specific detection of MC-LR in the range from 60 pM to 1000 nM with a detection limit of 15 pM. The credibility of the approach was confirmed by detection of MC-LR in real samples like serum and tap water samples. This study provides a new aptasensor for detection of MC-LR as well as other toxin analysis.

A rapid and simple ratiometric fluorescent sensor for patulin detection based on a stabilized DNA duplex probe containing less amount of aptamer-involved base pairs.

In this study, a sensor is described for determination of patulin by using ratiometric fluorescence measurement and strand displacement strategy. In the presence of patulin, the ratiometric fluorescence response decreases, owing to disassembly of DNA duplex structure and target-mediated release of TAMRA-labeled complementary DNA sequence2 (cDNA2). While, in the absence of target, the fluorescence resonance energy transfer (FRET) phenomenon happens between FAM and TAMRA under excitation at 490 nm, resulting in the enhancement of ratiometric signal. The use of ratiometric fluorescence signal with different signal indicators avoids the problem of environmental interference and improves the sensitivity of the aptasensor. Also, the DNA duplex structure contains minimum aptamer-involved base pair sequence, resulting in further improvement of the aptasensor sensitivity. This sensing platform provided a wide linear range from 15 ng/L to 35 µg/L and a detection limit of 6 ng/L for patulin. The aptasensor was used to determine patulin in spiked apple juice samples and showed satisfactory results.

Fluorescent sensor for detection of miR-141 based on target-induced fluorescence enhancement and PicoGreen.

Studies have shown that microRNAs affect the development of tumors. In many cases, they can be applied as biomarkers for the diagnosis of cancer; therefore, simple and sensitive analytical methods for detection of miRNAs are necessary. In this study, miR-141, which is used to diagnose several types of cancer, was detected in water and serum samples using a biosensor designed based on streptavidin-coated magnetic beads (SMBs), complementary sequences of miR-141 and PicoGreen as the fluorescent dye. The method is relatively fast and simple. Briefly, in the presence of miR-141, the complementary sequence forms a DNA:RNA double-strand on the surface of SMBs with intercalated PicoGreen. Upon attachment of the PicoGreen, the fluorescence intensity increased significantly (1000-fold). In the absence of a target, only single-stranded DNA (complementary strand of miR-141) existed on the surface of the SMBs. The fluorescence of the PicoGreen was low. The results revealed that the detection limits of the biosensor for miR-141 were 70 and 113.8 nmol L-1 in deionized water and serum samples, respectively.

Design, synthesis and biological evaluation of novel benzo- and tetrahydrobenzo-[h]quinoline derivatives as potential DNA-intercalating antitumor agents.

A new series of benzo- and tetrahydro benzo-[h]quinoline bearing a flexible (dimethylamino)ethylcarboxamide side chain was designed and synthesized as DNA-intercalating antitumor agents. The cytotoxic activity of the synthesized compounds was evaluated against four human cancer cell lines including MCF-7, A2780, C26 and A549. In general, saturated quinolines (tetrahydrobenzo[h]quinolines) exhibited more cytotoxicity compared to their corresponding unsaturated quinolines (benzo[h]quinolines). Compound 6e showed significant cytotoxicity against all four human cancer cell lines with IC50 values ranging from 1.86 to 3.91 µM. The interaction of the selected compounds showed significant cytotoxicity (6b, 6e, 6i and 6j) with calf thymus DNA (CT-DNA) was studied by UV and florescent spectroscopy. In general, benzo[h]quinolines showed higher interacting effect with DNA than their corresponding saturated tetrahydrobenzo[h]quinolines. Compound 6i exhibited the most DNA intercalating effects among the series. The apoptotic induction potential of the most cytotoxic compounds (6e, 6b and 6i) in A549 cells was studied using Annexin V-FITC/Propidium iodide staining assay. Compound 6e which showed the most cytotoxic effect against A549 cancer cells also exhibited stronger apoptotic induction activity in comparison with 6b and 6i.The docking was performed in order to study the DNA interaction properties of these compounds. According to the computational data, these compounds can interact with DNA as DNA-intercalating agents.

A label-free fluorescent aptasensor for detection of kanamycin based on dsDNA-capped mesoporous silica nanoparticles and Rhodamine B.

Kanamycin is an aminoglycoside antibiotic that can be useful against both gram negative and positive bacteria. However, if its serum levels are not controlled properly, it can cause serious side effects like ototoxicity and nephrotoxicity. The aim of this study was to design a simple and rapid fluorescent aptasensor for detection of kanamycin, based on Aptamer/Complementary strand (dsDNA)-capped mesoporous silica nanoparticles (MSNs) and Rhodamine B as a fluorescent probe. The MSNs pores were filled with Rhodamine B and then gated with dsDNA. In the presence of kanamycin, the aptamer sequence was separated from its complementary strand (CS), so that, uncovered the pores and leading to leakage of Rhodamine B. Thus, a significant increase in the fluorescence intensity was observed. The relative fluorescence intensity showed a linearity range from 24.75 nM to 137.15 nM of kanamycin with a detection limit of 7.5 nM. The aptasensor also showed to be useful for detection of kanamycin in serum samples and was able to distinguish kanamycin from other antibiotics, resulting in a sensitive, rapid and inexpensive method for kanamycin detection.

A colorimetric gold nanoparticle aggregation assay for malathion based on target-induced hairpin structure assembly of complementary strands of aptamer.

The authors describe a method for the colorimetric determination of the pesticide malathion. It is based on the use of a hairpin structure consisting of a complementary strand of aptamer and a double-stranded DNA (dsDNA) structure to protect gold nanoparticles (AuNPs) against salt-induced aggregation. In the absence of malathion, the dsDNA structure is preserved on the surface of AuNPs and the color of the AuNPs in solutions containing NaCl remains red. However, in the presence of malathion, a hairpin structure of complementary strand is formed. The Aptamer/Malathion complex and the complementary strand are released from the surface of the AuNPs. As a result, the AuNPs undergo salt-induced aggregation which is accompanied by a color change to blue. The assay allows malathion to be quantified within 35 min (A650/A520 was measured). The detection limit is 1 pM, and response is linear in the 5 pM to 10 nM malathion concentration range. The method is specific and was successfully applied to the determination of malathion in spiked human serum samples. Graphical abstract Schematic representation of detection of malathion based on dsDNA-modified gold nanoparticles (AuNPs) and the hairpin structure of the complementary strand.

Double targeting and aptamer-assisted controlled release delivery of epirubicin to cancer cells by aptamers-based dendrimer in vitro and in vivo.

Clinical use of epirubicin (Epi) in the treatment of cancer has been limited, due to its cardiotoxicity. Targeted delivery of chemotherapeutic agents could increase their efficacy and reduce their off-target effects. High drug loading and excellent stability of DNA dendrimers make these DNA nanostructures unique candidates for biological applications. In this study a modified and promoted dendrimer using three kinds of aptamers (MUC1, AS1411 and ATP aptamers) was designed for targeted delivery of Epi and its efficacy was evaluated in target cells including MCF-7 cells (breast cancer cell) and C26 cells (murine colon carcinoma cell). Aptamers (Apts)-Dendrimer-Epi complex formation was analyzed by fluorometric analysis and gel retardation assay. Release profiles of Epi from the designed complex were assessed at pHs 5.4 and 7.4. For MTT assay (cytotoxic study) MCF-7 and C26 cells (target cells) and CHO cells (Chinese hamster ovary cell, nontarget) were treated with Epi, Apts-Dendrimer-Epi complex and Apts-Dendrimer conjugate. Internalization was evaluated using flow cytometry analysis. Finally, the developed complex was used for inhibition of tumor growth in vivo. 25µM Epi was efficiently intercalated to 1µM dendrimer. Epi was released from the Apts-Dendrimer-Epi complex in a pH-sensitive manner (more release at pH 5.5). The results of flow cytometry analysis indicated that the designed complex was efficiently internalized into target cells, but not into control cells. The internalization data were confirmed by the results of MTT assay. Apts-Dendrimer-Epi complex had less cytotoxicity in CHO cells compared to Epi alone. The complex had more cytotoxicity in C26 and MCF-7 cells compared to Epi alone. Moreover, the Apts-Dendrimer-Epi complex could efficiently prohibit tumor growth in vivo. In conclusion, the designed targeted drug delivery system inherited characteristics of pH-dependent drug release, high drug loading and tumor targeting in vitro and in vivo.

Double targeting, controlled release and reversible delivery of daunorubicin to cancer cells by polyvalent aptamers-modified gold nanoparticles.

Clinical use of daunorubicin (Dau) in treatment of leukemia has been restricted because of its cardiotoxicity. Targeted delivery of anticancer drugs could decrease their off-target effects and enhance their efficacy. In this study a modified polyvalent aptamers (PA)-Daunorubicin (Dau)-Gold nanoparticles (AuNPs) complex was designed and its efficacy was assessed in Molt-4 cells (human acute lymphoblastic leukemia T-cell, target). Dau was efficiently loaded (10.5 µM) onto 1mL of PA-modified AuNPs. Dau was released from the PA-Dau-AuNPs complex in a pH-sensitive manner (faster release at pH5.5). The results of flow cytometry analysis indicated that the PA-Dau-AuNPs complex was efficiently internalized into target cells, but not into nontarget cells. The results of MTT assay were consistent with the internalization data. PA-Dau-AuNPs complex had less cytotoxicity in U266 cells compared to Dau alone and even Apt-Dau-AuNPs complex. The PA-Dau-AuNPs complex had more cytotoxicity in Molt-4 cells compared to Dau alone and even Apt-Dau-AuNPs complex. Cytotoxicity of PA-Dau-AuNPs complex was effectively antagonized using antisense of polyvalent aptamers. In conclusion, the designed drug delivery system inherited the properties of efficient drug loading, tumor targeting, pH-dependent drug release and controllable delivery of Dau to tumor cells.

Colorimetric and fluorescence quenching aptasensors for detection of streptomycin in blood serum and milk based on double-stranded DNA and gold nanoparticles.

Antibiotic residues in animal foodstuffs are of great concern to consumers. In this study, fluorescence quenching and colorimetric aptasensors were designed for detection of streptomycin based on aqueous gold nanoparticles (AuNPs) and double-stranded DNA (dsDNA). In the absence of streptomycin, aptamer/FAM-labeled complementary strand dsDNA is stable, resulting in the aggregation of AuNPs by salt and an obvious color change from red to blue and strong emission of fluorescence. In the presence of streptomycin, aptamer binds to its target and FAM-labeled complementary strand adsorbs on the surface of AuNPs. So the well-dispersed AuNPs remain stable against salt-induced aggregation with a wine-red color and the fluorescence of FAM-labeled complimentary strand is efficiently quenched by AuNPs. The colorimetric and fluorescence quenching aptasensors showed excellent selectivity toward streptomycin with limit of detections as low as 73.1 and 47.6 nM, respectively. The presented aptasensors were successfully used to detect streptomycin in milk and serum.

Synthesis, Characterization and Fluorescence Properties of Zn(II) and Cu(II) Complexes: DNA Binding Study of Zn(II) Complex.

Zinc(II) and copper(II) complexes containing Schiff base, 2- methoxy-6((E)-(phenylimino) methyl) phenol ligand (HL) were synthesized and characterized by elemental analysis, IR, NMR, and single crystal X-ray diffraction technique. The fluorescence properties and quantum yield of zinc complex were studied. Our data showed that Zn complex could bind to DNA grooves with Kb = 10(4) M(-1). Moreover, Zn complex could successfully be used in staining of DNA following agarose gel electrophoresis. MTT assay showed that Zn complex was not cytotoxic in MCF-7 cell line. Here, we introduce a newly synthesized fluorescence probe that can be used for single and double stranded DNA detection in both solution and agarose gels.

Targeted and controlled release delivery of daunorubicin to T-cell acute lymphoblastic leukemia by aptamer-modified gold nanoparticles.

Clinical administration of daunorubicin (Dau) in treatment of leukemia has been limited by its cardiotoxicity. Targeted delivery of chemotherapy drugs could reduce their side effects and increase the therapeutic efficacy of these drugs. Biocompatibility and large surface area of gold nanoparticles (AuNPs) make these nanoparticles great candidates for biomedical applications. In this study sgc8c aptamer (Apt)-Dau-AuNPs complex was designed and evaluated for treatment of Molt-4 cells (human acute lymphoblastic leukemia T-cell, target). Apt-Dau-AuNPs complex formation was analyzed by fluorometric analysis and gel retardation assay. Dau release profiles from the complex were evaluated in pHs 5.5 and 7.4. For cytotoxic studies (MTT assay) U266 (B lymphocyte human myeloma, nontarget) and Molt-4 cells (target) were treated with Dau Apt-Dau conjugate and Apt-Dau-AuNPs complex. Internalization was monitored by flow cytometry and confocal imaging. 12 µM Dau was efficiently loaded onto 1 mL of Apt-modified AuNPs. Dau was released from the complex in a pH-dependent manner (higher rate of release at pH 5.5). The results of flow cytometry analysis and confocal imaging showed that the complex was effectively internalized into Molt-4 cells, but not into U266 cells. The results of MTT assay also confirmed the internalization data. Apt-Dau-AuNPs complex was less cytotoxic in U266 cells compared to Dau alone and even Apt-Dau conjugate. The complex was more cytotoxic in target cells in comparison with Dau alone and even Apt-Dau conjugate. In conclusion, Apt-Dau-AuNPs complex was able to selectively target Molt-4 cells. Another advantage of this system was pH-dependent release of drug from the complex. Furthermore, this complex has characteristics which make it ideal for clinical use.

An aptasensor for selective, sensitive and fast detection of lead(II) based on polyethyleneimine and gold nanoparticles.

Lead (Pb), as a major environmental contaminant, could be harmful to humans when inhaled or ingested. In this study, we developed a sensitive, selective and fast colorimetric aptasensor for Pb(+2) based on polyethylenimine (PEI) and gold nanoparticles (AuNPs). In the absence of Pb(+2), aptamer binds to PEI. So the well-dispersed AuNPs remain stable with a wine-red color. Upon the addition of Pb(+2), a conformational change happens and a G-quadruplex aptamer/Pb(+2) complex is formed, leading to the aggregation of AuNPs and a color change to blue. This sensor showed a high selectivity toward Pb(+2) with a limit of detection (LOD) as low as 702pM. Moreover, our fabricated sensor was successfully applied for Pb(+2) detection in rat serum and tap water.

A novel colorimetric triple-helix molecular switch aptasensor for ultrasensitive detection of tetracycline.

Detection methods of antibiotic residues in blood serum and animal derived foods are of great interest. In this study a colorimetric aptasensor was designed for sensitive, selective and fast detection of tetracycline based on triple-helix molecular switch (THMS) and gold nanoparticles (AuNPs). As a biosensor, THMS shows distinct advantages including high stability, sensitivity and preserving the selectivity and affinity of the original aptamer. In the absence of tetracycline, THMS is stable, leading to the aggregation of AuNPs by salt and an obvious color change from red to blue. In the presence of tetracycline, aptamer binds to its target, signal transduction probe (STP) leaves the THMS and adsorbs on the surface of AuNPs. So the well-dispersed AuNPs remain stable against salt-induced aggregation with a red color. The presented aptasensor showed high selectivity toward tetracyclines with a limit of detection as low as 266 pM for tetracycline. The designed aptasensor was successfully applied to detect tetracycline in serum and milk.

Epirubicin loaded super paramagnetic iron oxide nanoparticle-aptamer bioconjugate for combined colon cancer therapy and imaging in vivo.

Every year a large number of new cases of colorectal cancer are diagnosed in the world. Application of Epirubicin (Epi) in treatment of cancer has been limited due to its cardiotoxicity. Specific delivery of chemotherapy drugs is an important factor in reducing the side effects of drugs used in chemotherapy. Enhanced permeability, retention effect and magnetic resonance (MR) traceability of super paramagnetic iron oxide nanoparticles (SPION) make them a great candidate in cancer therapy and imaging. In this study, Epirubicin-5TR1 aptamer-SPION tertiary complex was evaluated for the imaging and treatment of murine colon carcinoma cells (C26 cells, target). For cytotoxic studies (MTT assay), C26 and CHO-K1 (Chinese hamster ovary cells, nontarget) cells were treated with either Epi or Epi-Apt-SPION tertiary complex. Internalization was evaluated by flow cytometry. Finally, Apt-SPION bioconjugate was used for imaging of cancer in vivo. Flow cytometric analysis showed that the tertiary complex was internalized effectively to C26 cells, but not to CHO-K1 cells. Cytotoxicity of Epi-Apt-SPION tertiary complex also confirmed internalization data. The complex was less cytotoxic in CHO-K1 cells when compared to Epi alone. No significant change in viability between Epi- and complex-treated C26 cells was observed. Magnetic resonance imaging (MRI) indicated a high level of accumulation of the nano-magnets within the tumor site. In conclusion Epi-Apt-SPION tertiary complex is introduced as an effective system for targeted delivery of Epi to C26 cells. Moreover this complex could efficiently detect tumors when analyzed by MRI and inhibit tumor growth in vivo.

Reversible targeting and controlled release delivery of daunorubicin to cancer cells by aptamer-wrapped carbon nanotubes.

AIM: Single-walled carbon nanotubes (SWNTs) have been already used as drug carriers. In this study, we introduced sgc8c aptamer (this aptamer targets leukemia biomarker protein tyrosine kinase-7) to complex between Dau (daunorubicin) and SWNT to enhance targeted delivery of Dau to acute lymphoblastic leukemia T-cells (Molt-4). MATERIAL AND METHODS: Dau-aptamer-SWNTs tertiary complex formation was analyzed by visible spectroscopy and spectrofluorophotometric analysis. Dau release profiles from the complex were investigated in pH 7.4 and 5.5. For cytotoxic studies (MTT assay), Molt-4 (target) and U266 (B lymphocyte human myeloma, non-target) cells were treated with Dau, Dau-aptamer-SWNTs tertiary complex. Internalization was analyzed by flow cytometry. Targeted delivery of Dau was antagonized using antisense of aptamer. RESULTS: Dau was efficiently loaded onto SWNTs (efficiency ∼ 157%). Dau was released from Dau-aptamer-SWNTs tertiary complex in a pH-dependent manner (higher release rate at pH 5.5). Flow cytometric analysis showed that the tertiary complex was internalized effectively to Molt-4 cells, but not to U266 cells. Cytotoxicity of Dau-aptamer-SWNTs tertiary complex also confirmed internalization data. Dau-aptamer-SWNTs tertiary complex was less cytotoxic in U266 cells when compared to Dau alone. No significant change in viability between Dau- and complex-treated Molt-4 cells was observed. Cytotoxicity of Dau-aptamer-SWNTs complex was efficiently and quickly reversed using antisense in Molt-4 cells. CONCLUSION: Dau-aptamer-SWNTs complex is able to selectively target Molt-4 cells. The other advantages of this system are reversibility and pH-dependent release of Dau from its complex.