Screening of aptamers specific to colorectal cancer cells and stem cells by utilizing On-chip Cell-SELEX

Hung, Lien Yu; Wang, Chih‐Hung; Jui, Yu; Fu, Chien Yu; Chang, Hwan‐You; Wang, Kuan; Lee, Gwo‐Bin

doi:10.1038/srep10326

Cited by 55 publications

(33 citation statements)

References 47 publications

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“…CSC specific biomarkers are required for targeted delivery using aptamers. Researchers in the field have been focusing on improving the cell-SELEX protocol to select aptamers predominantly for CSCs [ 81 ]. Cell-surface markers such as the epithelial cell adhesion molecule (EpCAM), CD44, and CD133 have been tested for aptamer-mediated cell therapy [ 82 ].…”

Section: Recent Advances In Aptamer-sirna Applicationsmentioning

confidence: 99%

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Kruspe

Giangrande

2017

Biomedicines

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Synthetic nucleic acid ligands (aptamers) have emerged as effective delivery tools for many therapeutic oligonucleotide-based drugs, including small interfering RNAs (siRNAs). In this review, we summarize recent progress in the aptamer selection technology that has made possible the identification of cell-specific, cell-internalizing aptamers for the cell-targeted delivery of therapeutic oligonucleotides. In addition, we review the original, proof-of-concept aptamer-siRNA delivery studies and discuss recent advances in aptamer-siRNA conjugate designs for applications ranging from cancer therapy to the development of targeted antivirals. Challenges and prospects of aptamer-targeted siRNA drugs for clinical development are further highlighted.

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Section: Recent Advances In Aptamer-sirna Applicationsmentioning

confidence: 99%

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Kruspe

Giangrande

2017

Biomedicines

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“…In this regard, selection processes based on microfluidic chips have been proposed to render the SELEX more rapid and efficient [ 63 , 64 , 65 , 66 ]. These technologies have been mostly applied to protein-based selection and, only more recently, the Lee group reported a novel microfluidic system capable of performing automated cell-SELEX [ 67 , 68 ]. The microsystem used comprises three major modules: 1. a microfluidic control module, 2. a magnetic bead-based aptamer extraction module and 3. a temperature control system to regulate the reagent temperature and allow nucleic acid amplification by on-chip PCR.…”

Section: Cell-based Selex Methodsmentioning

confidence: 99%

“…Here, sequences are amplified for the subsequent round. The proposed approach was successfully used to isolate an aptamer specific for colorectal cancer cells and stem cells [ 67 ]. In addition, a similar strategy was applied to cholangiocarcinoma (CCA) cells, leading to isolation of three specific aptamers (one for SNU-478 cells and two for HuCCT-1 cells) [ 68 ].…”

Section: Cell-based Selex Methodsmentioning

confidence: 99%

Aptamer Cell-Based Selection: Overview and Advances

Catuogno

Esposito

2017

Biomedicines

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Aptamers are high affinity single-stranded DNA/RNA molecules, produced by a combinatorial procedure named SELEX (Systematic Evolution of Ligands by Exponential enrichment), that are emerging as promising diagnostic and therapeutic tools. Among selection strategies, procedures using living cells as complex targets (referred as “cell-SELEX”) have been developed as an effective mean to generate aptamers for heavily modified cell surface proteins, assuring the binding of the target in its native conformation. Here we give an up-to-date overview on cell-SELEX technology, discussing the most recent advances with a particular focus on cancer cell targeting. Examples of the different protocol applications and post-SELEX strategies will be briefly outlined.

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“…Images of antibody fluorescence were then captured using a DS-Qi1Mc camera (1.5 megapixels, equipped a Peltier cooling device and a programmable gain amplifier, Nikon) coupled to an inverted microscope equipped with a digital control module 31 . All optical and fluorescence images were captured using NIS-Elements Basic Research software (Br, version 4.20.00, 64 bit, Nikon, Japan).…”

Section: Methodsmentioning

confidence: 99%

A Microfluidic Chip for Detecting Cholangiocarcinoma Cells in Human Bile

Hung

Chiang

Tsai

et al. 2017

Sci Rep

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Cholangiocarcinoma (CCA), a biliary tract malignancy, accounts for 20% of all liver cancers. There are several existing methods for diagnosis of CCA, though they are generally expensive, laborious, and suffer from low detection rates. Herein we first developed a means of partially purifying human bile for consequent injection into a microfluidic chip. Then, the novel microfluidic system, which featured 1) a cell capture module, 2) an immunofluorescence (IF) staining module featuring two CCA-specific biomarkers, and 3) an optical detection module for visualization of antibody probes bound to these CCA marker proteins, was used to detect bile duct cancer cells within partially purified bile samples. As a proof of concept, CCA cells were successfully captured and identified from CCA cell cultures, blood samples inoculated with CCA cells, and clinical bile specimens. In 7.5 ml of bile, this system could detect >2, 0, and 1 positive cells in advanced stage patients, healthy patients, and chemotherapy-treated patients, respectively. In conclusion, our microfluidic system could be a promising tool for detection of cancer cells in bile, even at the earliest stages of CCA when cancer cells are at low densities relative to the total population of epithelial cells.

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Screening of aptamers specific to colorectal cancer cells and stem cells by utilizing On-chip Cell-SELEX

Cited by 55 publications

References 47 publications

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Aptamer Cell-Based Selection: Overview and Advances

A Microfluidic Chip for Detecting Cholangiocarcinoma Cells in Human Bile

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