Fluorescent signal aptamers are valuable molecular tools that can be used to establish important techniques or assay methods to determine the identity and concentration of proteins and metabolites. So far, Creative Biolabs has established a variety of methods to develop fluorescence modified aptamers.
Background of 3’ Inverted Thymidine Modified Aptamer
Aptamers are single-stranded DNA, RNA, and even modified nucleic acid molecules, which can form a clear tertiary structure and bind to specific targets. So far, a large number of artificial aptamers have been reported as affinity probes with high affinity and specificity for protein and small molecule metabolites. The availability of various aptamers has laid a solid foundation for exploring aptamers as highly coveted molecular tools for basic research and medical diagnosis.
However, in order to make full use of aptamers as affinity probes, it is necessary to find a way to connect the molecular recognition ability of aptamers with a simple signal generation process (no complex modification scheme is needed to change the aptamer to generate signals), versatility (regardless of what characteristics the aptamer and its target exhibit) and easy to use (only minimal sample manipulation is required in the signal acquisition process). Considering these factors, the combination of fluorescence technology and aptamers has become the preferred method.
Fig. 1 Chemical modification aptamers.1
Advantages of Fluorescence Modified Aptamer
Fluorescent aptamers have many attractive properties. In recent years, the design of aptamers with the fluorescence signal has become an important research activity.
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Fluorescence technology is highly compatible with aptamers. In the automatic synthesis process, many simple nucleic acids and fluorescent compounds can be well used in the synthesis process.
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The fluorescent group dressing of nucleic acid eliminates the need for target labeling, so it can be widely used in any aptamer-target pair.
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Fluorescence provides a very convenient way to report intermolecular interactions, because this detection can be performed in real-time without the need to separate the target-probe complex from the unbound probe.
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The availability of different fluorescent groups with different excitation and emission wavelengths makes it possible to perform multiple analyses.
Fluorescence Modified Aptamer Development in Creative Biolabs
The rational design strategies for converting aptamers into fluorescent dye probes can be summarized as: monomer method; dual chromophore method; duplex-to-complex structure-switching method; in-situ labeling method; chimeric aptamer method; dyeing method and aptamer-polymer conjugation method.
Creative Biolabs makes full use of our advantages and rich project experience to provide reasonable and customized design and development services of fluorescent modified aptamers for customers from all over the world. Please contact us to speak with a customer service representative.
References
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Ouellet, Jonathan. "RNA fluorescence with light-up aptamers." Frontiers in chemistry 4 (2016): 29.
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Questions & Answer
A: Commonly used fluorophores in aptamer modifications include FAM, Cy3, Cy5, and Alexa Fluor dyes. Cy3 and Cy5 are suitable for fluorescence resonance energy transfer (FRET) experiments, and Alexa Fluor dyes offer exceptional brightness and photostability. The choice of fluorophore depends on the specific application, such as imaging, biosensing, or flow cytometry.
A: Incorporating multiple fluorescence modifications can be challenging due to the potential for structural disruptions and decreased binding affinity. Researchers must carefully balance the number and placement of modifications to maintain aptamer functionality. Additionally, increased complexity can make synthesis and purification more demanding.
A: Recent advancements include the development of new fluorophores with improved properties, such as enhanced brightness and photostability. Additionally, researchers have been exploring advanced techniques like DNA nanotechnology to precisely position fluorophores within aptamer structures, allowing for better control over their effects on binding affinity and specificity.
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