The efficient cell-SELEX strategy, Icell-SELEX, using isogenic cell lines for selection and counter-selection to generate RNA aptamers to cell surface proteins

Takahashi, Masaki; Sakota, Eri; Nakamura, Yoshikazu

doi:10.1016/j.biochi.2016.09.018

Cited by 29 publications

(21 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Typically, cells exogenously overexpressing the target gene are used as the target cells, while the same wild type cells not expressing the target gene serve as negative cells [ 22 ]. Alternatively, negative cells can be engineered by silencing the expression of the target gene using RNA interference (RNAi) gene knockdown [ 23 ] or clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 gene editing [ 24 ]. Also, completely unrelated cells or non-malignant cells of the same tissue are commonly used for the counter-selection step [ 17 ].…”

Section: Aptamers As Ligands For the Targeted Delivery Of Therapeumentioning

confidence: 99%

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Kruspe

Giangrande

2017

Biomedicines

View full text Add to dashboard Cite

Synthetic nucleic acid ligands (aptamers) have emerged as effective delivery tools for many therapeutic oligonucleotide-based drugs, including small interfering RNAs (siRNAs). In this review, we summarize recent progress in the aptamer selection technology that has made possible the identification of cell-specific, cell-internalizing aptamers for the cell-targeted delivery of therapeutic oligonucleotides. In addition, we review the original, proof-of-concept aptamer-siRNA delivery studies and discuss recent advances in aptamer-siRNA conjugate designs for applications ranging from cancer therapy to the development of targeted antivirals. Challenges and prospects of aptamer-targeted siRNA drugs for clinical development are further highlighted.

show abstract

Section: Aptamers As Ligands For the Targeted Delivery Of Therapeumentioning

confidence: 99%

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Kruspe

Giangrande

2017

Biomedicines

View full text Add to dashboard Cite

show abstract

“…For counter selection, the same cell line was depleted in ITGAV with microRNA-mediated silencing. Takahashi et al [112] named this strategy Icell-SELEX for isogenic cell SELEX.…”

Section: Protein- and Cell-based Selex Processesmentioning

confidence: 99%

Selection of Nucleic Acid Aptamers Targeting Tumor Cell-Surface Protein Biomarkers

Mercier

Dontenwill

Choulier

2017

Cancers

View full text Add to dashboard Cite

Aptamers are nucleic acids referred to as chemical antibodies as they bind to their specific targets with high affinity and selectivity. They are selected via an iterative process known as ‘selective evolution of ligands by exponential enrichment’ (SELEX). Aptamers have been developed against numerous cancer targets and among them, many tumor cell-membrane protein biomarkers. The identification of aptamers targeting cell-surface proteins has mainly been performed by two different strategies: protein- and cell-based SELEX, when the targets used for selection were proteins and cells, respectively. This review aims to update the literature on aptamers targeting tumor cell surface protein biomarkers, highlighting potentials, pitfalls of protein- and cell-based selection processes and applications of such selected molecules. Aptamers as promising agents for diagnosis and therapeutic approaches in oncology are documented, as well as aptamers in clinical development.

show abstract

“…To overcome this problem, the same cell that is negative for desired antigen is used for counter SELEX. In current study transfected CD20 + HEK293T and HEK293T were used for SELEX and counter SELEX, respectively [ 25 , 26 ].…”

Section: Discussionmentioning

confidence: 99%

Selection and Characterization of Single-Stranded DNA Aptamers Binding Human B-Cell Surface Protein CD20 by Cell-SELEX

2018

View full text Add to dashboard Cite

The B-lymphocyte antigen (CD20) is a suitable target for single-stranded (ss) nucleic acid oligomer (aptamers). The aim of study was selection and characterization of a ssDNA aptamer against CD20 using Cell-Systematic Evolution of Ligands by Exponential Enrichment (Cell-SELEX). The cDNA clone of CD20 (pcDNA-CD20) was transfected to human embryonic kidney (HEK293T) cells. Ten rounds of Cell-SELEX was performed on recombinant HEK-CD20 cells. The final eluted ssDNA pool was amplified and ligated in T/A vector for cloning. The plasmids of positive clones were extracted, sequenced and the secondary structures of the aptamers predicted using DNAMAN® software. The sequencing results revealed 10 different types; three of them had the highest thermodynamic stability, named AP-1, AP-2 and AP-3. The AP-1 aptamer was the most thermodynamically stable one (ΔGAP-1 = −10.87 kcal/mol) with the highest binding affinity to CD20 (96.91 ± 4.5 nM). Since, the CD20 is a suitable target for recognition of B-Cell. The selected aptamers could be comparable to antibodies with many advantages. The AP-1, AP-2 and AP-3 could be candidate instead of antibodies for diagnostic and therapeutic applications in immune deficiency, autoimmune diseases, leukemia and lymphoma.

show abstract

The efficient cell-SELEX strategy, Icell-SELEX, using isogenic cell lines for selection and counter-selection to generate RNA aptamers to cell surface proteins

Cited by 29 publications

References 23 publications

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Aptamer-siRNA Chimeras: Discovery, Progress, and Future Prospects

Selection of Nucleic Acid Aptamers Targeting Tumor Cell-Surface Protein Biomarkers

Selection and Characterization of Single-Stranded DNA Aptamers Binding Human B-Cell Surface Protein CD20 by Cell-SELEX

Contact Info

Product

Resources

About