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Artificially Evolved Superbinder for Specific Recognition of N6-Methyladenine Base Modification in DNA and RNA.

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N6-Methyladenine (m6A/6mA) is a functional epigenetic base modification found in RNA and DNA. By selecting one RNA m6A reader as a template, we created a series of libraries of 3… Click to show full abstract

N6-Methyladenine (m6A/6mA) is a functional epigenetic base modification found in RNA and DNA. By selecting one RNA m6A reader as a template, we created a series of libraries of 3 × 108 RNA m6A reader mutants and developed a yeast surface-display-based evolution approach. Using high-throughput fluorescence-activated cell sorting, we ultimately obtained three evolved 6mA-binding proteins (e6mABPs), which displayed increased affinity for 6mA-containing DNA and reduced affinity for 6mA-free DNA. These e6mABPs are applicable for m6A/6mA enrichment and are potentially applied for modulating cell behavior.

Keywords: methyladenine; base modification; dna; rna

Journal Title: Analytical chemistry
Year Published: 2023

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