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Single-Stranded DNA Aptamers against Pathogens and Toxins: Identification and Biosensing Applications.

Hong KL, Sooter LJ - Biomed Res Int (2015)

Bottom Line: Molecular recognition elements (MREs) can be short sequences of single-stranded DNA, RNA, small peptides, or antibody fragments.There has been an increasing interest in the identification and application of nucleic acid molecular recognition elements, commonly known as aptamers, since they were first described in 1990 by the Gold and Szostak laboratories.Lastly, an overview of the basic principles of ssDNA aptamer-based biosensors is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Basic Pharmaceutical Sciences, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 20506, USA.

ABSTRACT
Molecular recognition elements (MREs) can be short sequences of single-stranded DNA, RNA, small peptides, or antibody fragments. They can bind to user-defined targets with high affinity and specificity. There has been an increasing interest in the identification and application of nucleic acid molecular recognition elements, commonly known as aptamers, since they were first described in 1990 by the Gold and Szostak laboratories. A large number of target specific nucleic acids MREs and their applications are currently in the literature. This review first describes the general methodologies used in identifying single-stranded DNA (ssDNA) aptamers. It then summarizes advancements in the identification and biosensing application of ssDNA aptamers specific for bacteria, viruses, their associated molecules, and selected chemical toxins. Lastly, an overview of the basic principles of ssDNA aptamer-based biosensors is discussed.

No MeSH data available.


Related in: MedlinePlus

Illustration of the SELEX process. A random library consisting of up to 1015 single-stranded nucleic acids molecules are incubated with the target of interest. Those that bound to the target are retrieved and amplified by polymerase case reaction. It is then followed by incubation with negative targets. Those that do not bind to negative targets are retained, amplified, and subjected to further rounds of in vitro selection.
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fig1: Illustration of the SELEX process. A random library consisting of up to 1015 single-stranded nucleic acids molecules are incubated with the target of interest. Those that bound to the target are retrieved and amplified by polymerase case reaction. It is then followed by incubation with negative targets. Those that do not bind to negative targets are retained, amplified, and subjected to further rounds of in vitro selection.

Mentions: In order to identify nucleic acid MREs, the SELEX process generally begins from a very large random library consisting of 1013 to 1015 different molecules. An individual nucleic acid MRE is composed of two constant regions for primer attachment during polymerase chain reaction (PCR) amplification flanked by 20–80 bases of random region [3]. The target of interest is first incubated with the library under specific ionic and temperature conditions. Library molecules that bind to the target are retained and amplified by PCR, while nonbinding library molecules are discarded. Negative or counter selections are often performed to increase the specificity of the library or direct the enrichment process away from binding to negative targets. Negative targets are often chosen for their structural similarities or the likelihood to coexist in the native environment with the target of interest. In this case, library molecules that bind to negative targets are discarded and those that do not bind are retained and amplified and thus completing one round of in vitro selection (Figure 1). It is expected that the library is enriched enough after approximately 12 rounds of SELEX. One or few nucleic acid MREs with high specificity and affinity toward their targets can be identified.


Single-Stranded DNA Aptamers against Pathogens and Toxins: Identification and Biosensing Applications.

Hong KL, Sooter LJ - Biomed Res Int (2015)

Illustration of the SELEX process. A random library consisting of up to 1015 single-stranded nucleic acids molecules are incubated with the target of interest. Those that bound to the target are retrieved and amplified by polymerase case reaction. It is then followed by incubation with negative targets. Those that do not bind to negative targets are retained, amplified, and subjected to further rounds of in vitro selection.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493287&req=5

fig1: Illustration of the SELEX process. A random library consisting of up to 1015 single-stranded nucleic acids molecules are incubated with the target of interest. Those that bound to the target are retrieved and amplified by polymerase case reaction. It is then followed by incubation with negative targets. Those that do not bind to negative targets are retained, amplified, and subjected to further rounds of in vitro selection.
Mentions: In order to identify nucleic acid MREs, the SELEX process generally begins from a very large random library consisting of 1013 to 1015 different molecules. An individual nucleic acid MRE is composed of two constant regions for primer attachment during polymerase chain reaction (PCR) amplification flanked by 20–80 bases of random region [3]. The target of interest is first incubated with the library under specific ionic and temperature conditions. Library molecules that bind to the target are retained and amplified by PCR, while nonbinding library molecules are discarded. Negative or counter selections are often performed to increase the specificity of the library or direct the enrichment process away from binding to negative targets. Negative targets are often chosen for their structural similarities or the likelihood to coexist in the native environment with the target of interest. In this case, library molecules that bind to negative targets are discarded and those that do not bind are retained and amplified and thus completing one round of in vitro selection (Figure 1). It is expected that the library is enriched enough after approximately 12 rounds of SELEX. One or few nucleic acid MREs with high specificity and affinity toward their targets can be identified.

Bottom Line: Molecular recognition elements (MREs) can be short sequences of single-stranded DNA, RNA, small peptides, or antibody fragments.There has been an increasing interest in the identification and application of nucleic acid molecular recognition elements, commonly known as aptamers, since they were first described in 1990 by the Gold and Szostak laboratories.Lastly, an overview of the basic principles of ssDNA aptamer-based biosensors is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Basic Pharmaceutical Sciences, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 20506, USA.

ABSTRACT
Molecular recognition elements (MREs) can be short sequences of single-stranded DNA, RNA, small peptides, or antibody fragments. They can bind to user-defined targets with high affinity and specificity. There has been an increasing interest in the identification and application of nucleic acid molecular recognition elements, commonly known as aptamers, since they were first described in 1990 by the Gold and Szostak laboratories. A large number of target specific nucleic acids MREs and their applications are currently in the literature. This review first describes the general methodologies used in identifying single-stranded DNA (ssDNA) aptamers. It then summarizes advancements in the identification and biosensing application of ssDNA aptamers specific for bacteria, viruses, their associated molecules, and selected chemical toxins. Lastly, an overview of the basic principles of ssDNA aptamer-based biosensors is discussed.

No MeSH data available.


Related in: MedlinePlus