Roman Teo Oliynyk

Reply to: Overestimations in the cyclization efficiency of small, restriction enzyme-digested DNA fragments

Roman Teo Oliynyk, George M. Church

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Short single-stranded DNA oligonucleotides enable intracellular transcription of functional RNAs in mammalian cells

Ahmed Mahas, Ramiro M. Perrotta, Roman Teo Oliynyk, Raphaël Ferreira et autres

Abstract Programmable intracellular production of short functional RNAs underlies diverse applications ranging from gene regulation to genome engineering, but is often constrained by cloning workflows, RNA synthesis, or viral-vector delivery. Here, we introduce a single-stranded DNA (ssDNA)-driven intracellular transcription strategy in which …

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DNA Cut-Ligation Cyclization Surpasses Jacobson–Stockmayer J-Factor Expectations by over Threefold

Roman Teo Oliynyk, George M. Church

For more than 75 years, the Jacobson-Stockmayer J-factor has been regarded as the fundamental physical limit on DNA cyclization efficiency. Here, we show that this limit can be substantially exceeded by simultaneous restriction cutting and ligation with the Type IIS enzyme BsaI-HFv2 …

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DNA Cut-Ligation Cyclization Surpasses Jacobson-Stockmayer J-Factor Expectations by Over Threefold

Roman Teo Oliynyk, George M. Church

Abstract We demonstrate DNA circularization efficiencies more than threefold greater than those predicted by classical Jacobson–Stockmayer theory for the ligation of linear double-stranded DNA. To quantify deviations from classical expectations, we experimentally calibrated the J-factor by ligating 452 bp DNA fragments bearing …

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Plasmid2MC: efficient cell-free generation of high-purity minicircle DNA for genome editing in mammalian cells

Roman Teo Oliynyk, Ahmed Mahas, Emil Karpinski

DNA plasmids are widely used for delivering proteins and RNA in genome editing. However, their bacterial components can lead to inactivation, cell toxicity, and reduced efficiency compared to minicircle DNA (mcDNA), which lacks such bacterial sequences. Existing commercial kits that recombine plasmids …

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Circular Vectors as an efficient, fully synthetic, cell-free approach for preparing small circular DNA as a plasmid substitute for guide RNA expression in CRISPR–Cas9 genome editing

Roman Teo Oliynyk, George M. Church

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Plasmid2MC: Efficient cell-free recombination of plasmids into high-purity minicircle DNA for use in genome editing applications

Roman Teo Oliynyk, Ahmed Mahas, Emil Karpinski

Abstract DNA plasmids are widely used for delivering proteins and RNA in genome editing. However, their bacterial components can lead to inactivation, cell toxicity, and reduced efficiency compared to minicircle DNA (mcDNA), which lacks such bacterial sequences. Existing commercial kits that recombine …

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Author Correction: Human-caused wolf mortality persists for years after discontinuation of hunting

Roman Teo Oliynyk

“The author is board member of the advocacy group Howling For Wolves. The position did not influence the work on this article and the advocacy group was not involved in design, analysis, or interpretation of the research.”

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Human-caused wolf mortality persists for years after discontinuation of hunting

Roman Teo Oliynyk

By the mid-twentieth century, wolves were nearly extinct in the lower 48 states, with a small number surviving in northern Minnesota. After wolves were placed on the endangered species list in 1973, the northern Minnesota wolf population increased and stabilized by the …

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Efficient modification and preparation of circular DNA for expression in cell culture

Roman Teo Oliynyk, George M. Church

DNA plasmids are an essential tool for delivery and expression of RNAs and proteins in cell culture experiments. The preparation of plasmids typically involves a laborious process of bacterial cloning, validation, and purification. While the expression plasmids can be designed and ordered …

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Human-caused wolf mortality persists for years after discontinuation of hunting

Roman Teo Oliynyk

Abstract By the mid-20th century, wolves were nearly extinct in the lower 48 states, with a small number surviving in northern Minnesota. After wolves were placed on the endangered species list in 1973, the northern Minnesota wolf population increased and stabilized by …

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Efficient Modification and Preparation of Circular DNA for Expression in Cell Culture

Roman Teo Oliynyk

Abstract DNA plasmids are an essential tool for delivery and expression of RNAs and proteins in cell culture experiments. The preparation of plasmids typically involves a laborious process of bacterial cloning, validation, and purification. While the expression plasmids can be designed and …

us, nz (code pays fourni par la source)