Generating Aptamers by Cell-SELEX for Applications in Molecular Medicine

Ye, Mao; Hu, Jun; Peng, Minyuan; Liu, Jing; Li, Jun; Liu, Huixia; Zhao, Xielan; Tan, Weihong

doi:10.3390/ijms13033341

Cited by 122 publications

(92 citation statements)

References 58 publications

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“…They are known to form conformationally unique 3-dimensional structures that allow for a selective interaction with their respective targets. Aptamers have been selected for a wide variety of targets ranging from small molecules to cells [1][2][3]. While aptamers are functionally similar to antibodies as molecular recognition elements, they do offer some advantages.…”

Section: Introductionmentioning

confidence: 99%

Aptamers as Promising Molecular Recognition Elements for Diagnostics and Therapeutics in the Central Nervous System

McConnell

Holahan

DeRosa

2014

Nucleic Acid Therapeutics

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Oligonucleotide aptamers are short, synthetic, single-stranded DNA or RNA able to recognize and bind to a multitude of targets ranging from small molecules to cells. Aptamers have emerged as valuable tools for fundamental research, clinical diagnosis, and therapy. Due to their small size, strong target affinity, lack of immunogenicity, and ease of chemical modification, aptamers are an attractive alternative to other molecular recognition elements, such as antibodies. Although it is a challenging environment, the central nervous system and related molecular targets present an exciting potential area for aptamer research. Aptamers hold promise for targeted drug delivery, diagnostics, and therapeutics. Here we review recent advances in aptamer research for neurotransmitter and neurotoxin targets, demyelinating disease and spinal cord injury, cerebrovascular disorders, pathologies related to protein aggregation (Alzheimer's, Parkinson's, and prions), brain cancer (glioblastomas and gliomas), and regulation of receptor function. Challenges and limitations posed by the blood brain barrier are described. Future perspectives for the application of aptamers to the central nervous system are also discussed.

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Section: Introductionmentioning

confidence: 99%

Aptamers as Promising Molecular Recognition Elements for Diagnostics and Therapeutics in the Central Nervous System

McConnell

Holahan

DeRosa

2014

Nucleic Acid Therapeutics

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“…The use of cell-SELEX has led to the generation of many aptamers for targeting live cells (25)(26)(27)(28)(29)(30)(31), and these aptamers have been found to be useful for the capture, detection, and imaging of cells, as well as for cancer imaging in vivo. However, practical applications of cancer-cell-specific aptamers are currently hampered by the lack of knowledge about their molecular targets, where only a very limited number of aptamer targets have been identified (7).…”

Section: Simultaneous Identifications Of Targets For Aptamersmentioning

confidence: 99%

“…Although recent studies have led to the selection of more than 100 cell aptamers, protein targets for only a very limited number of these aptamers have been identified (7), which greatly hampered their applications. In this vein, aptamer-target protein binding requires a native conformation of the aptamer.…”

mentioning

confidence: 99%

Facile Discovery of Cell-Surface Protein Targets of Cancer Cell Aptamers

Bing

Shangguan

Wang

2015

Molecular & Cellular Proteomics

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Cancer biomarker discovery constitutes a frontier in cancer research. In recent years, cell-binding aptamers have become useful molecular probes for biomarker discovery. However, there are few successful examples, and the critical barrier resides in the identification of the cellsurface protein targets for the aptamers, where only a limited number of aptamer targets have been identified so far. Herein, we developed a universal SILAC-based quantitative proteomic method for target discovery of cellbinding aptamers. The method allowed for distinguishing specific aptamer-binding proteins from nonspecific proteins based on abundance ratios of proteins bound to aptamer-carrying bait and control bait. In addition, we employed fluorescently labeled aptamers for monitoring and optimizing the binding conditions. We were able to identify and validate selectin L and integrin ␣4 as the protein targets for two previously reported aptamers, Sgc-3b and Sgc-4e, respectively. This strategy should be generally applicable for the discovery of protein targets for other cell-binding aptamers, which will promote the applications of these aptamers. Molecular & Cellular

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“…8,9 Both aptamers and DNAzymes are usually generated via Systematic Evolution of Ligands by Exponential enrichment (SELEX). [10][11][12][13] Many DNA and RNA aptamers have been derived to recognize a diverse range of targets including proteins, lipids, saccharides, nucleotides, metal ions, with high specificity and affinity. 14-23 DNA aptamers have also been selected to specifically recognize a host of tumor cells, such as human Burkitt's lymphoma cells, 24 glioblastoma cells, 25 liver cancer cells, 26 metastatic breast cancer cells, 27 acute leukemia cells, 28 brain tumor-initiating cells.…”

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confidence: 99%

Highly Specific Recognition of Breast Tumors by an RNA-Cleaving Fluorogenic DNAzyme Probe

Shen

et al. 2014

Anal. Chem.

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Breast cancer is one of the most commonly diagnosed cancers among females worldwide. Early detection of breast cancer is of vital importance to the reduction of the mortality rate. However, the lack of specific biomarkers that can effectively identify breast cancer cells limits the ability for early diagnosis of breast cancer. RNA-cleaving fluorogenic DNAzymes (RFDs), which can be produced through the systematic evolution of ligands by exponential enrichment (SELEX) process, are catalytic DNA molecules capable of generating a fluorescent signal when the appropriate target is bound. In this study, we carried out a SELEX experiment to select for RFDs that are active in the cell lysate of MDA-MB-231, a model breast cancer cell line. We obtained a RFD probe, named AAI2-5, that can detect MDA-MB-231 at a concentration of cell lysate proteins as low as 0.5 μg/mL (which is equivalent to ∼5000 cell/mL). AAI2-5 is capable of distinguishing MDA-MB-231 cells from normal cells as well as other types of tumor cells, including other subtypes of breast cancer cells. Moreover, AAI2-5 responded positively to more than 90% of malignant breast tumors. This report is the first study to explore the RFD system for the detection of cancer cells. The results suggest that RFD can be potentially applied for the diagnosis and treatment of breast cancer in the future.

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Generating Aptamers by Cell-SELEX for Applications in Molecular Medicine

Cited by 122 publications

References 58 publications

Aptamers as Promising Molecular Recognition Elements for Diagnostics and Therapeutics in the Central Nervous System

Aptamers as Promising Molecular Recognition Elements for Diagnostics and Therapeutics in the Central Nervous System

Facile Discovery of Cell-Surface Protein Targets of Cancer Cell Aptamers

Highly Specific Recognition of Breast Tumors by an RNA-Cleaving Fluorogenic DNAzyme Probe

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