Tag: pUC19

Development of Cas12a-Based Cell-Free Small-Molecule Biosensors via Allosteric Regulation of CRISPR Array Expression

In nature, microbes have evolved different systems to sense external stimuli. Synthetic biology approaches (1) repurpose these systems as biosensors to specifically and sensitively detect various targets of interest. Although various highly sensitive and specific laboratory-based analytical methods (including high-performance liquid chromatography and mass spectrometry) can detect small-molecule targets, they…

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Rapid, Efficient, and Cost-Effective Gene Editing of Enterococcus faecium with CRISPR-Cas12a

W. Hong et al. (12) previously demonstrated the utility of a CRISPR-Cas12a system in the low-GC Gram-positive organism C. difficile (12). As C. difficile and E. faecium have similar codon usage, we utilized Cas12a placed under the control of the tetracycline-inducible promoter (Ptet) from pRPF185 on the pMTL82151 backbone (pDL1,…

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Transformation of Lactobacillus acidophilus TK8912 by electroporation with pULA105E plasmid

An efficient method for the transformation of Lactobacillus acidophilus TK8912 by electroporation is presented. The plasmid vector pULA105E was constructed by joining a cryptic plasmid, pLA 105, from L. acidophilus TK8912, the Escherichia coli vector pUC19, and the erythromycin resistance gene of the Streptococcus-Escherichia coli shuttle vector pVA838. Plasmid pULA105E…

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High-purity production and precise editing of DNA base editing ribonucleoproteins

Abstract Ribonucleoprotein (RNP) complex–mediated base editing is expected to be greatly beneficial because of its reduced off-target effects compared to plasmid- or viral vector–mediated gene editing, especially in therapeutic applications. However, production of recombinant cytosine base editors (CBEs) or adenine base editors (ABEs) with ample yield and high purity in…

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