“…Multiplexing is especially beneficial with aptamer selections because it increases the likelihood of success per selection, where low success rates (30%) have been previously reported 9 23 . The SELEX process is also very time-consuming, often taking several weeks to months to complete 10 , so performing many selections simultaneously is extremely advantageous.…”
Section: Resultsmentioning
confidence: 99%
“…Because the conventional SELEX process is time consuming, many approaches have been proposed to decrease the time required for a selection 9 10 . RAPID-SELEX was previously developed by our group, which incorporates a single non-amplification cycle between cycles that include amplification of the enriched aptamer pool 11 .…”
Section: Resultsmentioning
confidence: 99%
“…The standard SELEX process tends to be time consuming and uses a large amount of reagents making aptamer selections costly 9 10 . To reduce time and reagent consumption, we developed a modified version of SELEX we called RNA APtamer Isolation via Dual-cycles (RAPID) 11 .…”
We describe a multiplexed RNA aptamer selection to 19 different targets simultaneously using a microcolumn-based device, MEDUSA (Microplate-based Enrichment Device Used for the Selection of Aptamers), as well as a modified selection process, that significantly reduce the time and reagents needed for selections. We exploited MEDUSA’s reconfigurable design between parallel and serially-connected microcolumns to enable the use of just 2 aliquots of starting library, and its 96-well microplate compatibility to enable the continued use of high-throughput techniques in downstream processes. Our modified selection protocol allowed us to perform the equivalent of a 10-cycle selection in the time it takes for 4 traditional selection cycles. Several aptamers were discovered with nanomolar dissociation constants. Furthermore, aptamers were identified that not only bound with high affinity, but also acted as inhibitors to significantly reduce the activity of their target protein, mouse decapping exoribonuclease (DXO). The aptamers resisted DXO’s exoribonuclease activity, and in studies monitoring DXO’s degradation of a 30-nucleotide substrate, less than 1 μM of aptamer demonstrated significant inhibition of DXO activity. This aptamer selection method using MEDUSA helps to overcome some of the major challenges with traditional aptamer selections, and provides a platform for high-throughput selections that lends itself to process automation.
“…Multiplexing is especially beneficial with aptamer selections because it increases the likelihood of success per selection, where low success rates (30%) have been previously reported 9 23 . The SELEX process is also very time-consuming, often taking several weeks to months to complete 10 , so performing many selections simultaneously is extremely advantageous.…”
Section: Resultsmentioning
confidence: 99%
“…Because the conventional SELEX process is time consuming, many approaches have been proposed to decrease the time required for a selection 9 10 . RAPID-SELEX was previously developed by our group, which incorporates a single non-amplification cycle between cycles that include amplification of the enriched aptamer pool 11 .…”
Section: Resultsmentioning
confidence: 99%
“…The standard SELEX process tends to be time consuming and uses a large amount of reagents making aptamer selections costly 9 10 . To reduce time and reagent consumption, we developed a modified version of SELEX we called RNA APtamer Isolation via Dual-cycles (RAPID) 11 .…”
We describe a multiplexed RNA aptamer selection to 19 different targets simultaneously using a microcolumn-based device, MEDUSA (Microplate-based Enrichment Device Used for the Selection of Aptamers), as well as a modified selection process, that significantly reduce the time and reagents needed for selections. We exploited MEDUSA’s reconfigurable design between parallel and serially-connected microcolumns to enable the use of just 2 aliquots of starting library, and its 96-well microplate compatibility to enable the continued use of high-throughput techniques in downstream processes. Our modified selection protocol allowed us to perform the equivalent of a 10-cycle selection in the time it takes for 4 traditional selection cycles. Several aptamers were discovered with nanomolar dissociation constants. Furthermore, aptamers were identified that not only bound with high affinity, but also acted as inhibitors to significantly reduce the activity of their target protein, mouse decapping exoribonuclease (DXO). The aptamers resisted DXO’s exoribonuclease activity, and in studies monitoring DXO’s degradation of a 30-nucleotide substrate, less than 1 μM of aptamer demonstrated significant inhibition of DXO activity. This aptamer selection method using MEDUSA helps to overcome some of the major challenges with traditional aptamer selections, and provides a platform for high-throughput selections that lends itself to process automation.
“…One particular approach to achieve acceptable aptamer stability is the use of Spiegelmer technology. For example, instead of endonuclease-sensitive RNA aptamers (D-form), researchers select endonuclease-resistant RNA aptamers (L-form) that result from binding to the mirror-image of the intended target molecule [173,174,175,176]. The mirror-image aptamer (L-form) subsequently is expected to bind the natural target molecule [173,177].…”
Section: Aptamers For Super-resolution Imagingmentioning
For many years, different probing techniques have mainly relied on antibodies for molecular recognition. However, with the discovery of aptamers, this has changed. The science community is currently considering using aptamers in molecular targeting studies because of the many potential advantages they have over traditional antibodies. Some of these possible advantages are their specificity, higher binding affinity, better target discrimination, minimized batch-to-batch variation, and reduced side effects. Overall, these characteristics of aptamers have attracted scholars to use them as molecular probes in place of antibodies, with some aptamer-based targeting products being now available in the market. The present review is aimed at discussing the potential of aptamers as probes in molecular biology and in super-resolution microscopy.
“…Compared with conventional SELEX methods, the developed microfluidic SELEX system dramatically decreased the incubation and partitioning time, thus simplifying the entire SELEX process. In addition, various newly developed separation and amplification technologies, including flow cytometry [ 86 , 87 ], biacore surface plasmon resonance [ 88 ], atomic force microscopy [ 89 – 91 ], and digital PCR [ 92 ] have been integrated into SELEX to obtain aptamers with high affinity and specificity to targets. Fig.…”
Section: Selection Of Specific Aptamers Using Selexmentioning
Aptamers are short non-coding, single-stranded oligonucleotides (RNA or DNA) developed through Systematic Evolution of Ligands by Exponential enrichment (SELEX) in vitro. Similar to antibodies, aptamers can bind to specific targets with high affinity, and are considered promising therapeutic agents as they have several advantages over antibodies, including high specificity, stability, and non-immunogenicity. Furthermore, aptamers can be produced at a low cost and easily modified, and are, therefore, called “chemical antibodies”. In the past years, a variety of aptamers specifically bound to both breast cancer biomarkers and cells had been selected. Besides, taking advantage of nanomaterials, there were a number of aptamer-nanomaterial conjugates been developed and widely investigated for diagnostics and targeted therapy of breast cancer. In this short review, we first present a systematical review of various aptamer selection methods. Then, various aptamer-based diagnostic and therapeutic strategies of breast cancer were provided. Finally, the current problems, challenges, and future perspectives in the field were thoroughly discussed.
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