CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch.
Thomsen EA, Skipper KA, Andersen S, Haslund D, Skov TW, Mikkelsen JG
Mol Ther Nucleic Acids. 2022 Aug 4;29:563-576. doi: 10.1016/j.omtn.2022.08.005. eCollection 2022 Sep 13.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 170540 | pT2/CMV-eGFP.PB/PTK | Expresses the puromycin resistance gene fused to a thymidine kinase, upon excision of this cassette the reading frame of eGFP is restored |
| 170542 | pPBT/EFS-Cas9-PuroTk | Expresses spCas9 with a P2A junction to the puromycin resistance gene directly fused to the thymidine kinase |
| 170543 | pLV/Guide-mCherry | Expresses mCherry and contains a cassette for U6 driven sgRNA transcription |
| 170544 | pLV/eGFP-U6-sgRNA | A lentiviral backbone for homology directed insertion of eGFP. Containing only eGFP, flanked by restriction sites for insertion of homology arms and a sgRNA casstte to clone in sgRNA for HDR |
| 170545 | pLV.5’-LMNA-eGFP-KI | Expresses a sgRNA for 5' tagging to LMNA and contains a cassette with eGFP flanked by homology arms for LMNA |
| 170546 | pLV.5’eGFP/sgLMNA | Expresses a sgRNA for 5' tagging to LMNA and contains a cassette with eGFP without homology arms |
| 170547 | pLV.5’VIM-eGFP-KI | Expresses a sgRNA for 5' tagging to VIM and contains a cassette with eGFP flanked by homology arms for VIM |
| 170548 | pLV.5’eGFP/sgVIM | Expresses a sgRNA for 5' tagging to VIM and contains a cassette with eGFP without homology arms |