Targeted DNA integration in human cells without double-strand breaks using CRISPR-associated transposases.
Lampe GD, King RT, Halpin-Healy TS, Klompe SE, Hogan MI, Vo PLH, Tang S, Chavez A, Sternberg SH
Nat Biotechnol. 2023 Mar 29. doi: 10.1038/s41587-023-01748-1. PubMed Article
Lampe GD, King RT, Halpin-Healy TS, Klompe SE, Hogan MI, Vo PLH, Tang S, Chavez A, Sternberg SH
Nat Biotechnol. 2023 Mar 29. doi: 10.1038/s41587-023-01748-1. PubMed Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 200892 | pSL2620 (pTniQ_Vch) | Expresses human codon-optimized V. cholerae TniQ from a CMV promoter. |
| 200893 | pSL2621 (pCas8_Vch) | Expresses human codon-optimized V. cholerae Cas8 from a CMV promoter. |
| 200894 | pSL2622 (pCas7_Vch) | Expresses human codon-optimized V. cholerae Cas7 from a CMV promoter. |
| 200895 | pSL2623 (pCas6_Vch) | Expresses human codon-optimized V. cholerae Cas6 from a CMV promoter. |
| 200896 | pSL2783 (pTnsC-VP64_Vch) | Expresses human codon-optimized V. cholerae TnsC-VP64 from a CMV promoter. |
| 200897 | pSL2669 (pTnsABf_Vch) | Expresses human codon-optimized V. cholerae TnsA-TnsB fusion from a CMV promoter. |
| 200898 | pSL0812 (pCRISPR(BbsI)_Vch) | Expresses V. cholerae CRISPR RNA from a U6 promoter. The CRISPR array contains two BbsI sites for spacer cloning. |
| 200899 | pSL3718 (pQCascade_Pse) | Expresses human codon-optimized Pseudoalteromonas Cas7, Cas8, Cas6, and TniQ from a CMV promoter, in a polycistronic vector format separated by T2A sequences. |
| 200900 | pSL3713 (pTnsC_Pse) | Expresses human codon-optimized Pseudoalteromonas TnsC from a CMV promoter. |
| 200901 | pSL3402 (pTnsABf_Pse) | Expresses human codon-optimized Pseudoalteromonas TnsA-TnsB fusion from a CMV promoter. |
| 200902 | pSL3617 (pCRISPR(BbsI)_Pse) | Expresses Pseudoalteromonas CRISPR RNA from a U6 promoter. The CRISPR array contains two BbsI sites for spacer cloning. |
| 200903 | pSL3936 (pDonor(AAVS1)_Pse) | Encodes a mini-transposon derived from Pseudoalteromonas Tn7016 with a SA-T2A-PuroR-T2A-eGFP cargo. Total transposon size = 2,024 bp. |
| 200904 | pSL5069 (pDonor(AAVS1),CRISPR_Pse) | Encodes a mini-transposon derived from Pseudoalteromonas Tn7016 with a splice acceptor-T2A-PuroR-T2A-eGFP cargo, and a CRISPR RNA under a U6 promoter. Total transposon size = 2,024 bp. |
| 200905 | pSL3927 (pDonor(plasmid)_Pse) | Encodes a mini-transposon derived from Pseudoalteromonas Tn7016,with a CmrR cargo, as well as a primer binding site for deep sequencing purposes at pTarget. Total transposon size = 1,087 bp. |
| 200906 | pSL0341 (pTarget) | Encodes an mCherry casette under a minimal CMV promoter. Used as the target plasmid for plasmid-based integration assays, and for plasmid-based transcriptional activation. |
| 200907 | pSL4393 (pClpX_Eco) | Expresses human codon-optimized E. coli ClpX from a CMV promoter. |