Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: PAM-interacting domain swapping is extensively utilized in nature to evolve CRISPR-Cas9 nucleases with altered PAM specificities It has been anticipated that protospacer adjacent motif (PAM) specificity of the CRISPR-Cas9 nucleases protospacer appears to be modular. Here we present the finding that naturally occurring domain swapping has been extensively involved in altering its PAM specificities. Sequence analysis of streptococcal Cas9 sequences revealed conservation of three distinct PAM-interacting motifs, with phylogenetic analysis of full-length Cas9 and the PID demonstrating that PAM-interacting domain (PID) domain swapping was extensively utilized to diversify its PAM specificity. An extended analysis of 582 representative Cas9 sequences revealed that this PID swapping was broadly present in most of the investigated genera. Mimicking the natural PID domain swapping, a functional chimeric enzyme, based on the scaffold of compact Staphylococcus aureus Cas9, with novel NNAAAA PAM specificity was developed. In summary, our findings shed new light on a rich source of exchangeable PID domains in Cas9 genes, which can be mined for domain swapping aiming to an effective PAM refinement.
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The categories of this paper are: systems
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The categories of this paper are: plant
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The categories of this paper are: ecology
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The categories of this paper are: synthetic
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The categories of this paper are: developmental
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The categories of this paper are: immunology
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The categories of this paper are: cognition
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The categories of this paper are: epidemiology
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The categories of this paper are: pathology
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The categories of this paper are: and
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The categories of this paper are: biophysics
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The categories of this paper are: neuroscience
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The categories of this paper are: microbiology
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The categories of this paper are: toxicology
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The categories of this paper are: bioengineering
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The categories of this paper are: bioinformatics