scholarly journals Development of aptamer-based inhibitors for CRISPR/Cas system

2020 ◽  
Author(s):  
Jing Zhao ◽  
Rika Inomata ◽  
Yoshio Kato ◽  
Makoto Miyagishi
Keyword(s):  
Genome Editing ◽  
In Vitro Selection ◽  
Unsolved Problem ◽  
Dna Aptamer ◽  
Cleavage Assay ◽  
Locked Nucleic Acids ◽  
Systematic Evolution ◽  
Exponential Enrichment ◽  
In Cells

Abstract The occurrence of accidental mutations or deletions caused by genome editing with CRISPR/Cas9 system remains a critical unsolved problem of the technology. Blocking excess or prolonged Cas9 activity in cells is considered as one means of solving this problem. Here, we report the development of an inhibitory DNA aptamer against Cas9 by means of in vitro selection (systematic evolution of ligands by exponential enrichment) and subsequent screening with an in vitro cleavage assay. The inhibitory aptamer could bind to Cas9 at low nanomolar affinity and partially form a duplex with CRISPR RNA, contributing to its inhibitory activity. We also demonstrated that improving the inhibitory aptamer with locked nucleic acids efficiently suppressed Cas9-directed genome editing in cells and reduced off-target genome editing. The findings presented here might enable the development of safer and controllable genome editing for biomedical research and gene therapy.

Aptamer Oligonucleotides as Potential Therapeutics in Hematologic Diseases

2019 ◽  
Vol 19 (10) ◽  
pp. 788-795
Author(s):  
Weibin Li ◽  
Meng Zhao ◽  
Huihui Yan ◽  
Kaiyu Wang ◽  
XIaopeng lan
Keyword(s):  
Clinical Trials ◽  
In Vitro Selection ◽  
Cell Disease ◽  
Anemia Of Chronic Disease ◽  
The Past ◽  
Systematic Evolution ◽  
Anemia Of Chronic Inflammation ◽  
Exponential Enrichment ◽  
Potential Therapeutics

: Aptamers are single-stranded DNA or RNA oligonucleotides generated by a novel in vitro selection technique termed Systematic evolution of ligands by exponential enrichment (SELEX). During the past two decades, various aptamer drugs have been developed and many of them have entered into clinical trials. : In the present review, we focus on aptamers as potential therapeutics for hematological diseases, including anemia of chronic inflammation (ACI) and anemia of chronic disease (ACD), hemophilia, thrombotic thrombocytopenic purpura (TTP) or VWD type-2B, and sickle cell disease (SCD), in particular, those that have entered into clinical trials

scholarly journals In vitro selection of an RNA aptamer yields an interleukin-6/interleukin-6 receptor interaction inhibitor

2021 ◽  
Author(s):  
Takehiro Ando ◽  
Mizuki Yamamoto ◽  
Yukio Takamori ◽  
Keita Tsukamoto ◽  
Daisuke Fuji ◽  
...  
Keyword(s):  
Interleukin 6 ◽  
In Vitro Selection ◽  
Receptor Interaction ◽  
Rna Aptamer ◽  
Cytokine Storm ◽  
Interleukin 6 Receptor ◽  
Systematic Evolution ◽  
Exponential Enrichment ◽  
Selection Of

ABSTRACT Interleukin-6 (IL-6) binds to IL-6 receptor (IL-6R) subunit, related to autoimmune diseases and cytokine storm in COVID-19. In this study we performed Systematic Evolution of Ligands by Exponential enrichment (SELEX) and identified a novel RNA aptamer. This RNA aptamer not only bound to IL-6R with a dissociation constant of 200 nM, but also inhibited the interaction of IL-6R with IL-6.

scholarly journals Back Cover: In Vitro Selection of a DNA Aptamer Targeting Degraded Protein Fragments for Biosensing (Angew. Chem. Int. Ed. 20/2020)

2020 ◽  
Vol 59 (20) ◽  
pp. 7968-7968
Author(s):  
Meng Liu ◽  
Jiayi Wang ◽  
Yangyang Chang ◽  
Qiang Zhang ◽  
Dingran Chang ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Dna Aptamer ◽  
Protein Fragments ◽  
Back Cover ◽  
Selection Of

Differentiating breast cancer molecular subtypes using a DNA aptamer selected against MCF-7 cells

2018 ◽  
Vol 6 (12) ◽  
pp. 3152-3159 ◽  
Author(s):  
Mei Liu ◽  
Tong Yang ◽  
Zhongsi Chen ◽  
Zhifei Wang ◽  
Nongyue He
Keyword(s):  
Breast Cancer ◽  
Molecular Subtypes ◽  
Personalized Therapy ◽  
Dna Aptamer ◽  
Single Stranded Dna ◽  
Systematic Evolution ◽  
Exponential Enrichment ◽  
Mcf 7

Aptamers are single-stranded DNA or RNA oligonucleotides selected by systematic evolution of ligands by exponential enrichment (SELEX), which show great potential in the diagnosis and personalized therapy of cancers, due to their specific advantages over antibodies.

scholarly journals Implementation of High-Throughput Sequencing (HTS) in Aptamer Selection Technology

2020 ◽  
Vol 21 (22) ◽  
pp. 8774
Author(s):  
Natalia Komarova ◽  
Daria Barkova ◽  
Alexander Kuznetsov
Keyword(s):  
Nucleic Acid ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Combinatorial Libraries ◽  
Structure And Function ◽  
Huge Number ◽  
Systematic Evolution ◽  
And Function ◽  
Exponential Enrichment

Aptamers are nucleic acid ligands that bind specifically to a target of interest. Aptamers have gained in popularity due to their high potential for different applications in analysis, diagnostics, and therapeutics. The procedure called systematic evolution of ligands by exponential enrichment (SELEX) is used for aptamer isolation from large nucleic acid combinatorial libraries. The huge number of unique sequences implemented in the in vitro evolution in the SELEX process imposes the necessity of performing extensive sequencing of the selected nucleic acid pools. High-throughput sequencing (HTS) meets this demand of SELEX. Analysis of the data obtained from sequencing of the libraries produced during and after aptamer isolation provides an informative basis for precise aptamer identification and for examining the structure and function of nucleic acid ligands. This review discusses the technical aspects and the potential of the integration of HTS with SELEX.

In Vitro Selection of a DNA Aptamer Targeting Degraded Protein Fragments for Biosensing

2020 ◽  
Vol 59 (20) ◽  
pp. 7706-7710 ◽  
Author(s):  
Meng Liu ◽  
Jiayi Wang ◽  
Yangyang Chang ◽  
Qiang Zhang ◽  
Dingran Chang ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Dna Aptamer ◽  
Protein Fragments ◽  
Selection Of

scholarly journals Aptamers selection for platelets using droplet digital PCR

2021 ◽  
pp. 100-103
Author(s):  
A.V. Blagodatova ◽  
◽  
K.V. Kochkina ◽  
M.A. Komarova ◽  
N.Y. Trofina ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Anticoagulant Activity ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Platelet Glycoprotein ◽  
Systematic Evolution ◽  
Selection For ◽  
Exponential Enrichment ◽  
Selection Of

The aim of the research. To obtain aptamers-inhibitors of platelet glycoprotein IIb / IIIa receptors, blocking platelet aggregation. Material and methods. Th e selection of aptamers for IIb / IIIa receptors of platelets was carried out according to the SELEX method (Systematic Evolution of Ligands by Exponential Enrichment), modifi ed to select aptamers for a specifi c epitope. Th e method allows selection and in vitro evolution of aptamers with selectivity to a specifi c target from a large library of oligonucleotides. Th e affi nity of aptamers for platelet IIb / IIIa receptors was determined using fl ow cytometry. Results. Pools of aptamers of aptamers with high affi nity for IIb / IIIa platelet receptors were obtained. Th e study of the antiaggregation properties of the pools with the best binding showed that platelet aggregation was minimal when using the aptamers from the pool of the 5th round of selection. Th us, the aptamers of this pool have the greatest potential to be used as an analogue of a synthetic peptide that blocks thromboaggregation. Aptamers from this pool were taken for sequencing in order to obtain sequences of aptamers with the best antiaggregatory properties. Conclusion. Pools of aptamers with high affi nity for IIb / IIIa receptors of platelets and anticoagulant activity were obtained.

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