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DNA cut-ligation cyclization surpasses J-Factor limit by order of magnitude

Oliynyk, R. T.; Church, G. M.

2026-01-31 molecular biology
10.64898/2026.01.31.702778 bioRxiv
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We demonstrate DNA circularization efficiencies more than threefold greater than those predicted by classical Jacobson-Stockmayer theory - a cornerstone physical model unchallenged for over seven decades - even at high DNA concentrations suitable for routine practical use. This breakthrough performance is enabled by simultaneous restriction cutting and ligation of double-stranded DNA fragments using the Type IIS enzyme BsaI-HFv2 in combination with T4 DNA ligase under finely optimized buffer conditions. These results unveil a biologically derived exception that overcomes the longstanding physical barriers to DNA cyclization imposed by Jacobson-Stockmayer theory, while highlighting the potential for systematic exploration of enzyme combinations to identify additional systems achieving comparable or superior intramolecular ligation efficiency.

Published in Biomolecules (predicted rank #3) · training set

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