Products/Services Used | Details | Operation |
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GenParts™ DNA Fragments> | … DNA fragments were synthesized by Genscript while oligonucleotides were synthesized by IDT (Integrated DNA Technologies, USA). … The whole cassette was synthesized by Genscript and assembled into the pCDF-1b plasmid (ColDF13 ori, Millipore, USA) replacing the MCS … | Get A Quote |
CRISPR base editing is a powerful method to engineer bacterial genomes. However, it restricts editing to single-nucleotide substitutions. Here, to address this challenge, we adapt a CRISPR-Prime Editing-based, DSB-free, versatile, and single-nucleotide resolution genetic manipulation toolkit for prokaryotes. It can introduce substitutions, deletions, insertions, and the combination thereof, both in plasmids and the chromosome of E. coli with high fidelity. Notably, under optimal conditions, the efficiency of 1-bp deletions reach up to 40%. Moreover, deletions of up to 97 bp and insertions up to 33 bp were successful with the toolkit in E. coli, however, efficiencies dropped sharply with increased fragment ... More