TY - JOUR T1 - CRISPR/Cas9-Assisted Transformation-Efficient Reaction (CRATER), a novel method for selective transformation JF - bioRxiv DO - 10.1101/027664 SP - 027664 AU - L. J. Rothschild AU - D. T. Greenberg AU - J. R. Takahashi AU - K. A. Thompson AU - A. J. Maheshwari AU - R. E. Kent AU - G. McCutcheon AU - J. D. Shih AU - C. Calvet AU - T. D. Devlin AU - T. Ju AU - D. Kunin AU - E. Lieberman AU - T. Nguyen AU - F. G. Tran AU - D. Xiang AU - K. Fujishima Y1 - 2015/01/01 UR - http://biorxiv.org/content/early/2015/09/27/027664.abstract N2 - The CRISPR/Cas9 system has revolutionized genome editing by providing unprecedented DNA-targeting specificity. Here we demonstrate that this system can be applied to facilitate efficient plasmid selection for transformation as well as selective gene insertion into plasmid vectors by cleaving unwanted plasmid byproducts after restriction enzyme digestion and ligation. Using fluorescent and chromogenic proteins as reporters, we demonstrate that CRISPR/Cas9 cleavage excludes unwanted ligation byproducts and increases transformation efficiency of desired inserts from 20% up to 97% ± 3%. This CRISPR/Cas9-Assisted Transformation-Efficient Reaction (CRATER) protocol is a novel, inexpensive, and convenient method for obtaining specific cloning products. ER -