Precise DNA cleavage using CRISPR-SpRYgests.
Christie KA, Guo JA, Silverstein RA, Doll RM, Mabuchi M, Stutzman HE, Lin J, Ma L, Walton RT, Pinello L, Robb GB, Kleinstiver BP
Nat Biotechnol. 2022 Oct 6. pii: 10.1038/s41587-022-01492-y. doi: 10.1038/s41587-022-01492-y.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 181743 | pET28-SpRY-NLS-6xHis (gbr2101) | bacterial expression plasmid for SpRY with an n-terminal MKIEE MBP tag, and c-terminal bi-partite NLS and 6x-His tag |
| 181744 | pET28-SpRY-HF1-NLS-6xHis (gbr2102) | bacterial expression plasmid for SpRY-HF1 with an n-terminal MKIEE MBP tag, and c-terminal bi-partite NLS and 6x-His tag |
| 181745 | pACYC-T7-SpCas9-T7-EGFP_gRNA1-T7term (BPK848) | dual T7 promoter vector for bacterial expression of SpCas9 with a c-terminal NLS and 3xFLAG tag, along with an SpCas9 sgRNA targeting EGFP site 1 |
| 181746 | pTox-EGFPsite1_NGG-PAM (BPK740) | p11-lacY-wtx1 derivative toxic plasmid encoding an SpCas9 target site for EGFP site 1 with an NGG PAM |
| 181747 | pTox-EGFPsite1_NGA-PAM (BPK754) | p11-lacY-wtx1 derivative toxic plasmid encoding an SpCas9 target site for EGFP site 1 with an NGA PAM |
| 181748 | pUC19-SpRYgest_targetplasmid (KAC833) | pUC19 derivative plasmid used as a substrate for SpRYgest, restriction enzyme, and TtAgo digests. Encodes an SpCas9 EMX1-site1-NGG-PAM target site between NheI and HindIII restriction sites. |