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Click chemistry is a versatile chemical reaction which allows for the synthesis of complex conjugates when direct biomolecule conjugation falls short. The click reaction uses bio-orthogonal or biologically unique functional groups to join two distinct molecules in a two-step process. Bio-Synthesis can introduce these functional group such as azide or alkyne to oligonucleotides, peptides or any biomolecule during chemical synthesis or post-synthetically.
All custom modified DNA, RNA oligos, peptide to be use for click chemistry are produced at our state-of-the-art oligo facility in Lewisville Texas. Each modified oligo is meticulously monitored during synthesis and controlled according to Bio-Synthesis' stringent ISO 9001 quality standards. The final product is inspected by mass spectrometry and its purity is analyzed by high throughput gel electrophoresis.
Contact Bio-Synthesis for oligonucleotide or peptide modifications, labeling and conjugation for click chemistry.
There are several organic reactions that fit the definition of click chemistry. For modification of oligonucleotides, we offer oligonucleotide modification for copper catalyzed click chemistry reaction. We also offer oligonucleotide modifications or conjguates using copper-free click chemistry and many other bioorthogonal probes. See information on Bioorthogonal Probes.
Click chemistry was demonstrated by Sharpless and co-workers in 2001 to describe the [3+2] cycloaddition using copper-catalyzed triazole formation of an azide and alkyne. The two components - azido and alkyne groups - are nearly never encountered in natural biomolecules. Hence, conjugation reactions arrive at a remarkable degree of selectivity and specificity. The development of the click methodology has led to the production and study of many modified DNA click chemistry probes for biological as well as nanotechnological and surface oriented applications.
Azide and alkynes click reaction are sterospecific, inert, stable and small in size. These two functional groups are interchangeable; either one can tag your target molecule, while the other is use for subsequent detection. The label is small enough that tagged molecules are acceptable substrates for enzymes that assemble biopolymers. The reaction can be conducted in benign, easily removable solvents, and side products can be easily removed by non chromatographic methods. As result, this chemistry often used to generate very large combinatorial libraries of small-molecule compounds to be screen as drug candidates. Click chemistry offers the potential to accelerate the drug discovery process, as it makes each reaction in the multi-step synthesis of a small molecule fast, efficient and predictable. Click Chemistry technology is reliable and stable which makes it an ideal oligonucleotide labeling method.
The reaction between an azide and an alkyne has been referred to as the Huisgen cycloaddition reaction since 1964. More recently the Sharpless group invented the term “click chemistry” to describe the reactions because of the nature of the electrophiles that participate in it.
Azides are useful electrophilic contributors in click chemistry reactions due to their ease of formation and stability. For this reason, alkyl azides undergo nearly no side reactions and are extremely stable in aqueous solution regardless of the presence of biological material.
In addition, alkyne groups are also remarkably stable in biological mixtures, as long as they don’t have an activating group nearby, such as a carbonyl, which would make them susceptible to Michael-type addition reactions, especially with thiols.
Both functional groups used in click chemistry conjugations are completely unreactive towards biological molecules and free from side reactions which would cause instability in aqueous environments. As a result, oligonucleotides modified to contain an azide or alkyne group would react specifically to one another even in the presence of other molecules. In addition, without the presence of Cu(I), the azido-molecule and the alkyne-molecule would not by catalyzed to react to an considerable extent.
There is one drawback to the classic click chemistry approach. The copper ions used in the reaction can harm cells, reduce fluorescence of fluorophores and even impair protein functions. An alternative for preventing these harmful reactions is to modify an oligo with DCBO alkyne reagents. DCBO modified oligonucleotides are compatible with copper-free click chemistry reactions.
Copper(I)-catalyzed alkyne-azide [3+2] cycloaddition click reaction offers a great potential for synthesis of unique oligo based research tools. Examples include conjugating cell penetrating peptides (CPP) to antisense or siRNA to increase cell uptake or labeling of an oligonucleotide with biotin and/or fluorescent dyes in multiple sites within same oligo have routinely been performed at Bio-Synthesis.
Applications of Click chemistry have a vigorous effect on the following areas:
Bio-Synthesis offers ready-to-use modified custom oligonucleotide synthesis to be used for click chemistry reactions. As always, Bio-Synthesis strives to be at the forefront of technical improvements in order to serve our customers best. We offer a collection of alkynes modified oligonucleotide synthesis for the classic CuAAC click chemistry reaction, as well as the streamlined copper-free click modifications and other bioorthogonal probes.
To order an alkyne- or azide-modified oligonucleotide:specify the nucleic acid sequence along with the appropriate alkyne or azide modifications included in the sequence.
Browse our selection of modified bases using the tabs below. When you ready to design and order your oligonucleotide, simply click on the order button. Alternatively, let us design the oligonucleotide for you.