Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA integration.
Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH
Nature. 2019 Jul;571(7764):219-225. doi: 10.1038/s41586-019-1323-z. Epub 2019 Jun 12.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 130633 | pSL0283 (pTnsABC) | Expresses V. cholerae CAST TnsA, TnsB, and TnsC from a T7 promoter. |
| 130634 | pSL0527 (pDonor) | Encodes a mini-transposon derived from V. cholerae CAST system, with CmR cargo gene. Total transposon size = 977 bp. |
| 130635 | pSL0284 (pQCascade_entry) | Expresses V. cholerae CAST TniQ, Cas8, Cas7, and Cas6 from one T7 promoter, and a CRISPR RNA from a second T7 promoter. The CRISPR array contains two BsaI sites for spacer cloning. |
| 130636 | pSL0762 (pQCascade_crRNA-nt) | Expresses V. cholerae CAST TniQ, Cas8, Cas7, and Cas6 from one T7 promoter, and a CRISPR RNA from a second T7 promoter. The CRISPR array encodes a non-targeting guide RNA. |
| 130637 | pSL0828 (pQCascade_crRNA-4) | Expresses V. cholerae CAST TniQ, Cas8, Cas7, and Cas6 from one T7 promoter, and a CRISPR RNA from a second T7 promoter. The CRISPR array encodes a guide RNA that targets lacZ. |
| 130638 | pSL0373 (His-MBP-TniQ) | Expresses V. cholerae CAST TniQ from a T7 promoter, with N-terminal His10-MBP-TEVsite tag for purification. |
| 130639 | pSL0374 (His-MBP-Cas8_Cas7_Cas6) | Expresses V. cholerae CAST Cas8, Cas7, and Cas6 from a T7 promoter, with N-terminal His10-MBP-TEVsite tag on Cas8 for purification. |
| 130640 | pSL0375 (His-MBP-TniQ_Cas8_Cas7_Cas6) | Expresses V. cholerae CAST TniQ, Cas8, Cas7, and Cas6 from a T7 promoter, with N-terminal His10-MBP-TEVsite tag on TniQ for purification. |
| 130641 | pSL0376 (His-MBP-StrepII-TniQ) | Expresses V. cholerae CAST TniQ from a T7 promoter, with N-terminal His10-MBP-TEVsite-StrepII for purification. |
| 130642 | pSL0426 (CRISPR with spc3) | Expresses CRISPR RNA from a T7 promoter for VchCAST system. The CRISPR array encodes a guide RNA with spacer 3 from the native CRISPR. |
| 130643 | pSL0915 (CRISPR with target 3) | Expresses CRISPR RNA from a T7 promoter for VchCAST system. The CRISPR array encodes a guide RNA that targets lacZ. |
| 130644 | pSL0715 (pDonor_R1-57) | Encodes a mini-transposon derived from V. cholerae Tn6677 CAST, with CmR cargo gene and shortened right end. Total transposon size = 910 bp. |
| 130645 | pSL0721 (pDonor_L1-105) | Encodes a mini-transposon derived from V. cholerae Tn6677 CAST, with CmR cargo gene and shortened left end. Total transposon size = 933 bp. |
| 130646 | pSL0746 (pDonor_R-MmeI) | Encodes a mini-transposon derived from V. cholerae Tn6677 CAST, with CmR cargo gene. The mutated right end introduces an MmeI recognition site (used for Tn-seq). Total transposon size = 977 bp. |
| 130647 | pSL0747 (pDonor_L-MmeI) | Encodes a mini-transposon derived from V. cholerae Tn6677 CAST, with CmR cargo gene. The mutated left end introduces an MmeI recognition site (used for Tn-seq). Total transposon size = 977 bp. |