The clustered regularly interspaced short palindrome repeats (CRISPR) and Crispr-associated protein 9 (CRISPR/Cas9) is now a well-known, revolutionary method to engineer microbial cells.
A key advantage of CRISPR remains in the strain design to facilitate chromosomal integration to enable the assembly of marker-free DNA. These editing systems are highly beneficial; however, their assembly is not quite straightforward and may prevent its use and applications., Tigran V. Yuzbashev and a research team identified the limits of the existing Cas9 toolkits designed to make CRISPR techniques easier to access and implement.
A single toolkit comprised of 147 plasmids to generate and characterize a library of 137 promoters to build aThe CRISPR/Cas9 system can render quick, precise and scarless genomic modifications to provide significant scope to design microbial strains for bioproduction.
To enable CRISPR-based marker-free integration, the team chose a double strand break induced by Cas9, which had to be repaired to accomplish cell proliferation. The scientists made this possible by using a template or donor, integrated through
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