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Development of DNA aptamers using Cell-SELEX.

Kwame Sefah¹, Dihua Shangguan, Xiangling Xiong

¹Center for Research at Bio/Nano Interface, Department of Chemistry, University of Florida, Gainesville, Florida, USA.

Nature Protocols

|June 12, 2010

Summary

This study details cell-SELEX, a method to create DNA aptamers that bind specifically to target cells, like cancer cells. This technique enriches aptamer pools for precise cell targeting and differentiation.

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Area of Science:

Biotechnology
Molecular Biology
Genomics

Background:

Nucleic acid aptamers are highly specific binding molecules.
Aptamers can be generated for diverse targets, including live cells.
Cancer cell-specific aptamers are valuable for diagnostics and therapeutics.

Purpose of the Study:

To describe the cell-SELEX protocol for generating DNA aptamers against specific cell types.
To enable the differentiation of target cells (e.g., cancer cells) from other cell types.

Main Methods:

The cell-SELEX protocol involves iterative steps of cell binding, washing, recovery, and amplification.
DNA aptamer pools are incubated with target cells, and non-binders are removed.
Selection against control cells removes non-specific binders, enriching for target-specific aptamers.

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Polymerase chain reaction (PCR) amplifies bound aptamers, and antisense strands are removed for subsequent rounds.

Flow cytometry assays monitor aptamer pool enrichment through increased fluorescence.

Main Results:

The cell-SELEX method successfully enriches DNA aptamer pools with high specificity for target cells.
Selected aptamers demonstrate increased binding affinity and specificity compared to the unselected library.
The protocol allows for the generation of aptamers capable of distinguishing between different cell types.

Conclusions:

Cell-SELEX is an effective method for developing DNA aptamers for specific cell targeting.
This protocol provides a powerful tool for isolating aptamers against complex biological targets like cancer cells.
The generated aptamers have potential applications in cell-based diagnostics and targeted therapies.