CRISPR/Cpf1-Mediated Multiplex and Large-Fragment Gene Editing in Staphylococcus aureus

Staphylococcus aureus is
a major
human pathogen that causes a variety of infections, including life-threatening
diseases. Research on S. aureus is
constrained by complex and limited genetic manipulation methods. Here,
we report a CRISPR/Cpf1-mediated system, pCpfSA, for rapid and versatile
genome editing in S. aureus. In direct
comparison with the existing CRISPR/Cas9-mediated genome-editing system,
the pCpfSA system exhibits enhanced colony-forming units (CFUs) after
editing and an expanded targetable range with comparable editing efficiency.
Given the precursor crRNA (pre-crRNA) processing activity of Cpf1,
the pCpfSA system also allows multiplex gene editing and large-fragment
DNA knockout simply by introducing two crRNAs and the corresponding
donor templates, which is difficult to achieve using the CRISPR/Cas9
system, thereby greatly expanding the genome editor toolbox for S. aureus.