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Development of multiplexed orthogonal base editor (MOBE) systems

Nat Biotechnol. 2024-05; 
Quinn T Cowan, Sifeng Gu, Wanjun Gu, Brodie L Ranzau, Tatum S Simonson, Alexis C Komor
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Codon Optimization … Codon-optimized (GenScript) CPs were ordered as a gBlock gene fragment (Integrated DNA technologies) and subcloned into CP-deaminase fusions with uracil-specific excision … Get A Quote

摘要

Base editors (BEs) enable efficient, programmable installation of point mutations while avoiding the use of double-strand breaks. Simultaneous application of two or more different BEs, such as an adenine BE (which converts A·T base pairs to G·C) and a cytosine BE (which converts C·G base pairs to T·A), is not feasible because guide RNA crosstalk results in non-orthogonal editing, with all BEs modifying all target loci. Here we engineer both adenine BEs and cytosine BEs that can be orthogonally multiplexed by using RNA aptamer-coat protein systems to recruit the DNA-modifying enzymes directly to the guide RNAs. We generate four multiplexed orthogonal BE systems that enable rates of precise co-occurring edits... More

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