In Vitro and Ex Vivo Selection Procedures for Identifying Potentially Therapeutic DNA and RNA Molecules

Marton, Soledad; Reyes-Darias, José Antonio; Sánchez‐Luque, Francisco J.; Romero-López, Cristina; Berzal‐Herranz, Alfredo

doi:10.3390/molecules15074610

Cited by 26 publications

(31 citation statements)

References 167 publications

(179 reference statements)

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“…2d). Just as with DNA-based aptamers 126 , it is possible to create a population of structures with variable segments for downstream selection according to desired function or even overall structure. The development of methods to evolve single-stranded DNA structures is undoubtedly a challenge worth pursuing.…”

Section: Technical Challengesmentioning

confidence: 99%

Challenges and opportunities for structural DNA nanotechnology

et al. 2011

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DNA molecules have been used to build a variety of nanoscale structures and devices over the past 30 years, and potential applications have begun to emerge. But the development of more advanced structures and applications will require a number of issues to be addressed, the most significant of which are the high cost of DNA and the high error rate of self-assembly. Here we examine the technical challenges in the field of structural DNA nanotechnology and outline some of the promising applications that could be developed if these hurdles can be overcome. In particular, we highlight the potential use of DNA nanostructures in molecular and cellular biophysics, as biomimetic systems, in energy transfer and photonics, and in diagnostics and therapeutics for human health.

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Section: Technical Challengesmentioning

confidence: 99%

Challenges and opportunities for structural DNA nanotechnology

et al. 2011

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“…During the selection step, a subpopulation of individual sequences specifically interacts with the target molecule and this enriched subpopulation is isolated by physicochemical techniques [11,12,13]. Subsequently, bound sequences are eluted and amplified for a next round of selection in which the stringency of the selection conditions may be increased to identify the tightest-binding sequences.…”

Section: Aptamers: a Potential Diagnostic And Therapeutic Alternativementioning

confidence: 99%

“…Individual aptamers can be further modified in order to improve their approachability [12,15]. For instance, chemical modifications may be introduced rising the aptamer in vivo stability.…”

Section: Aptamers: a Potential Diagnostic And Therapeutic Alternativementioning

confidence: 99%

Use of Aptamers as Diagnostics Tools and Antiviral Agents for Human Viruses

González

Martı́n

Fernández³

et al. 2016

Pharmaceuticals

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Appropriate diagnosis is the key factor for treatment of viral diseases. Time is the most important factor in rapidly developing and epidemiologically dangerous diseases, such as influenza, Ebola and SARS. Chronic viral diseases such as HIV-1 or HCV are asymptomatic or oligosymptomatic and the therapeutic success mainly depends on early detection of the infective agent. Over the last years, aptamer technology has been used in a wide range of diagnostic and therapeutic applications and, concretely, several strategies are currently being explored using aptamers against virus proteins. From a diagnostics point of view, aptamers are being designed as a bio-recognition element in diagnostic systems to detect viral proteins either in the blood (serum or plasma) or into infected cells. Another potential use of aptamers is for therapeutics of viral infections, interfering in the interaction between the virus and the host using aptamers targeting host-cell matrix receptors, or attacking the virus intracellularly, targeting proteins implicated in the viral replication cycle. In this paper, we review how aptamers working against viral proteins are discovered, with a focus on recent advances that improve the aptamers’ properties as a real tool for viral infection detection and treatment.

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“…Accordingly, aptamers are reproduced by chemical synthesis, their smaller size allows more efficient entry into biological compartments, and they are non-immunogenic. In addition, aptamers are easily labeled with fluorescent or other reporters during their synthesis [16], they can be chemically modified to become extremely stable [17,18], or can be further truncated to eliminate oligonucleotide sequences which are not important for the interaction with the target or for the correct tridimensional aptamer structure [19,20]. Consequently, they are considered potential diagnostic and therapeutic agents and also thought to be rival antibodies in immunoassay-like analyses.…”

Section: Introductionmentioning

confidence: 99%

DNA Aptamers Selectively Target Leishmania infantum H2A Protein

et al. 2013

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Parasites of the genus Leishmania produce leishmaniasis which affects millions people around the world. Understanding the molecular characteristics of the parasite can increase the knowledge about the mechanisms underlying disease development and progression. Thus, the study of the molecular features of histones has been considered of particular interest because Leishmania does not condense the chromatin during mitosis and, consequently, a different role for these proteins in the biology of the parasite can be expected. Furthermore, the sequence divergences in the amino and in the carboxy-terminal domains of the kinetoplastid core histones convert them in potential diagnostic and/or therapeutics targets. Aptamers are oligonucleotide ligands that are selected in vitro by their affinity and specificity for the target as a consequence of the particular tertiary structure that they are able to acquire depending on their sequence. Development of high-affinity molecules with the ability to recognize specifically Leishmania histones is essential for the progress of this kind of study. Two aptamers which specifically recognize Leishmania infantum H2A histone were cloned from a previously obtained ssDNA enriched population. These aptamers were sequenced and subjected to an in silico analysis. ELONA, slot blot and Western blot were performed to establish aptamer affinity and specificity for LiH2A histone and ELONA assays using peptides corresponding to overlapped sequences of LiH2A were made mapping the aptamers:LiH2A interaction. As “proofs of concept”, aptamers were used to determine the number of parasites in an ELONA platform and to purify LiH2A from complex mixtures. The aptamers showed different secondary structures among them; however, both of them were able to recognize the same peptides located in a side of the protein. In addition, we demonstrate that these aptamers are useful for LiH2A identification and also may be of potential application as diagnostic system and as a laboratory tool with purification purpose.

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In Vitro and Ex Vivo Selection Procedures for Identifying Potentially Therapeutic DNA and RNA Molecules

Cited by 26 publications

References 167 publications

Challenges and opportunities for structural DNA nanotechnology

Challenges and opportunities for structural DNA nanotechnology

Use of Aptamers as Diagnostics Tools and Antiviral Agents for Human Viruses

DNA Aptamers Selectively Target Leishmania infantum H2A Protein

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