Response to Comment on “RNA-guided DNA insertion with CRISPR-associated transposases”
Jonathan Strecker, Alim Ladha, Kira S. Makarova, Eugene V. Koonin, Feng Zhang
Abstract
Rice et al . suggest that the CRISPR-associated transposase ShCAST system could lead to additional insertion products beyond simple integration of the donor. We clarify the outcomes of ShCAST-mediated insertions in Escherichia coli , which consist of both simple insertions and integration of the donor plasmid. This latter outcome can be avoided by use of a 5′ nicked DNA donor.
Topics & Concepts
TransposaseCRISPRPlasmidEscherichia coliDNABiologyInsertion sequenceDNA Transposable ElementsComputational biologyGeneticsTransposable elementGeneGenomeCRISPR and Genetic EngineeringBacterial Genetics and BiotechnologyRNA and protein synthesis mechanisms