Abstract
A chemical ligation method for construction of DNA-encoded small-molecule libraries has been developed. Taking advantage of the ability of the Klenow fragment of DNA polymerase to accept templates with triazole linkages in place of phosphodiesters, we have designed a strategy for chemically ligating oligonucleotide tags using cycloaddition chemistry. We have utilized this strategy in the construction and selection of a small molecule library, and successfully identified inhibitors of the enzyme soluble epoxide hydrolase.
Highlights
DNA-recording approaches require the iterative construction of both the chemical library members and the encoding oligonucleotide
Previous work had qualitatively shown that triazole-containing oligonucleotides could be amplified by PCR and that a triazole-containing plasmid could be translated in living bacteria[17,18] or human cells[19]; direct quantitation of read-through efficiency was not reported
Oligonucleotide 3 was synthesized using a CuAAC ligation followed by biotin labeling of the 5′ -amine linker (Fig. 1)
Summary
Alexander Litovchick[1], Christoph E. Taking advantage of the ability of the Klenow fragment of DNA polymerase to accept templates with triazole linkages in place of phosphodiesters, we have designed a strategy for chemically ligating oligonucleotide tags using cycloaddition chemistry We have utilized this strategy in the construction and selection of a small molecule library, and successfully identified inhibitors of the enzyme soluble epoxide hydrolase. Recent reports from Brown, El-Sagheer and Tavassolli have demonstrated that oligonucleotides containing a triazole linkage in place of a phosphodiester are competent substrates for PCR and could provide a “readable” encoding sequence[16,17,18,19] Based on these results, and our prior experience with Cu-catalyzed alkyne-azide cycloaddition (CuAAC) of oligos[20], we wondered whether a readable chemical ligation strategy might offer some advantages over the current enzymatic methods. We wondered if the 20-base sequence upstream of the triazole could be reduced in length while
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