PAM-Flexible Genome Editing with an Engineered Chimeric Cas9.
Journal
Research square
Titre abrégé: Res Sq
Pays: United States
ID NLM: 101768035
Informations de publication
Date de publication:
07 Mar 2023
07 Mar 2023
Historique:
entrez:
22
3
2023
pubmed:
23
3
2023
medline:
23
3
2023
Statut:
epublish
Résumé
CRISPR enzymes require a defined protospacer adjacent motif (PAM) flanking a guide RNA-programmed target site, limiting their sequence accessibility for robust genome editing applications. In this study, we recombine the PAM-interacting domain of SpRY, a broad-targeting Cas9 possessing an NRN > NYN PAM preference, with the N-terminus of Sc++, a Cas9 with simultaneously broad, efficient, and accurate NNG editing capabilities, to generate a chimeric enzyme with highly flexible PAM preference: SpRYc. We demonstrate that SpRYc leverages properties of both enzymes to specifically edit diverse NNN PAMs and disease-related loci for potential therapeutic applications. In total, the unique approaches to generate SpRYc, coupled with its robust flexibility, highlight the power of integrative protein design for Cas9 engineering and motivate downstream editing applications that require precise genomic positioning.
Identifiants
pubmed: 36945419
doi: 10.21203/rs.3.rs-2625838/v1
pmc: PMC10029082
pii:
doi:
Types de publication
Preprint
Langues
eng
Subventions
Organisme : NCI NIH HHS
ID : DP2 CA281401
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL142494
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM115911
Pays : United States
Commentaires et corrections
Type : UpdateIn
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