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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. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
200892pSL2620 (pTniQ_Vch)Expresses human codon-optimized V. cholerae TniQ from a CMV promoter.
200893pSL2621 (pCas8_Vch)Expresses human codon-optimized V. cholerae Cas8 from a CMV promoter.
200894pSL2622 (pCas7_Vch)Expresses human codon-optimized V. cholerae Cas7 from a CMV promoter.
200895pSL2623 (pCas6_Vch)Expresses human codon-optimized V. cholerae Cas6 from a CMV promoter.
200896pSL2783 (pTnsC-VP64_Vch)Expresses human codon-optimized V. cholerae TnsC-VP64 from a CMV promoter.
200897pSL2669 (pTnsABf_Vch)Expresses human codon-optimized V. cholerae TnsA-TnsB fusion from a CMV promoter.
200898pSL0812 (pCRISPR(BbsI)_Vch)Expresses V. cholerae CRISPR RNA from a U6 promoter. The CRISPR array contains two BbsI sites for spacer cloning.
200899pSL3718 (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.
200900pSL3713 (pTnsC_Pse)Expresses human codon-optimized Pseudoalteromonas TnsC from a CMV promoter.
200901pSL3402 (pTnsABf_Pse)Expresses human codon-optimized Pseudoalteromonas TnsA-TnsB fusion from a CMV promoter.
200902pSL3617 (pCRISPR(BbsI)_Pse)Expresses Pseudoalteromonas CRISPR RNA from a U6 promoter. The CRISPR array contains two BbsI sites for spacer cloning.
200903pSL3936 (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.
200904pSL5069 (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.
200905pSL3927 (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.
200906pSL0341 (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.
200907pSL4393 (pClpX_Eco)Expresses human codon-optimized E. coli ClpX from a CMV promoter.

Antibodies from Article