What is SELEX method?

What is SELEX method?

SELEX is an experimental procedure that involves the progressive selection, from a large combinatorial double-stranded oligonucleotide library, of DNA ligands with variable DNA-binding affinities and specificities by repeated rounds of partition and amplification.

How aptamers can be designed and produced?

Aptamer is an artificial chemical antibody that is generated from the randomized nucleic acid library by three simple steps: binding, separation, and amplification. The selected aptamer has a high binding affinity with the target molecule. In most cases, aptamer was found to be better than antibodies.

How are aptamers produced?

Aptamers are generated via an in vitro process known as the Systematic Evolution of Ligands by EXponential enrichment, also known as SELEX. Tuerk and Gold (1990), aiming to find an RNA aptamer sequence that would bind T4 DNA polymerase, first termed the process.

What is SELEX what can it be used for?

Systematic Evolution of Ligands by EXponential Enrichment (SELEX) is a common method currently used for isolating high-affinity single-stranded (ss) DNAs or RNAs from a large library with random sequences [1,2,3].

How do aptamers work?

It works by isolating sequences with increased affinity and specificity to a target molecule from an oligo sequence pool through several rounds of selection. DNA or RNA molecules that bind to the target are prospective aptamers for that target.

What is counter selection SELEX?

As selection has evolved, researchers have gone beyond negative selection for generic SELEX components and added negative selection (counter selection) steps that actively seek to remove sequences binding to unwanted targets from the oligonucleotide pool.

How aptamers bind to their targets?

Aptamers are nucleic acid macromolecules that bind to molecular targets, including proteins, with high affinity and specificity. Stable tertiary structure, resulting from combinations of these secondary structures, allows aptamers to bind to targets via van der Waals, hydrogen bonding, and electrostatic interactions.

What does the aptamer sequence do?

Aptamers are single-stranded oligonucleotides that fold into defined architectures and bind to targets such as proteins. In binding proteins they often inhibit protein–protein interactions and thereby may elicit therapeutic effects such as antagonism.

What is RNA aptamers?

RNA Aptamers refer to RNA oligonulceotides that are capable of binding to specific targets with high affinity and specificity. RNA aptamers have proven to be of high therapeutic and diagnostic value with recent FDA approval of the first aptamer drug and additional ones in the clinical pipelines.

How do you take SELEX?

Specifically, SELEX process includes (1) the incubation of target molecules with the random sequence pools, (2) and the subsequent separation of unbound oligonucleotides and the elution of bound oligonucleotides, (3) then PCR amplification of bound aptamers.

Who invented SELEX?

Professor Larry Gold, and then graduate student Craig Tuerk developed the SELEX process. A randomized pool of around 65,500 nucleotide sequences underwent the process, and two of these sequences were isolated. The experiment’s results led to the concept that nucleic acids could be used to recognize proteins.

What is SELEX aptamer?

SELEX Aptamer Selection. Aptamers with affinity for a desired target are selected from a large oligonucleotide library through a process called SELEX, which stands for Sequential Evolution of Ligands by Exponential Enrichment. This improves the selectivity of the resulting aptamer candidates.

How are aptamers used in RNA therapeutics?

Both RNA and DNA aptamers are being developed for various uses, including therapeutics [1]. The high-affinity binding of aptamers is accomplished by folding the aptamer in a three-dimensional structure specific to the target protein. Aptamers were first introduced in 1990 by two independent research groups [2,3].

Are there any aptamers that target blood coagulation?

Although no aptamers targeting blood coagulation factors have been approved for clinical use, several aptamers have been evaluated in clinical trials and many more have demonstrated encouraging preclinical results.

Which is the best aptamer for protein binding?

Aptamers are single-stranded nucleic acids, either DNA or RNA, that bind to their target protein with high affinity and specificity. During the early years of aptamers technology, RNA aptamers were preferred since RNA was assumed to fold in a more functional domain. However, recently, DNA aptamers are becoming more prevalent [1].

Why are aptamer oligonucleotides used in bio sensing?

The specificity of the aptamers and other unique attributes such as ease in chemical synthesis, tunable backbone modification, low immunogenicity, and excellent stability [5,6], allow aptamer oligonucleotides to offer multiple advantages over antibodies, in medical diagnostic, bio-sensing, and bio-imaging platforms.

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