Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.
Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, Kleinstiver BP
Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x.
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Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 208942 | Click editor (CE1) - pCMV-T7-PCV2-nCas9-EcKlenow (EMK488) | The CE1 construct, expressed from CMV or T7 promoters. |
| 208943 | Control click editor - pCMV-T7-deadPCV2-nCas9-EcKlenow (EMK492) | A control CE1 construct with catalytically attenuated PCV2 HUH, expressed from CMV or T7 promoters. |
| 208944 | Control click editor - pCMV-T7-PCV2-deadCas9-EcKlenow (EMK500) | A control CE1 construct with catalytically inactivated SpCas9, expressed from CMV or T7 promoters. |
| 208945 | Control click editor - pCMV-T7-PCV2-nCas9-deadEcKlenow (JO520) | A control CE1 construct with catalytically attenuated EcKlenow DNA polymerase, expressed from CMV or T7 promoters. |
| 208946 | HUHe variant click editor (CE1) - pCMV-T7-DCV-nCas9-EcKlenow (JO608) | A variant CE1 construct with DCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 208947 | HUHe variant click editor (CE1) - pCMV-T7-MSMV-nCas9-EcKlenow (JO661) | A variant CE1 construct with MSMV HUH endonuclease, expressed from CMV or T7 promoters. |
| 208948 | HUHe variant click editor (CE1) - pCMV-T7-TraI-nCas9-EcKlenow (JO604) | A variant CE1 construct with TraI HUH endonuclease, expressed from CMV or T7 promoters. |
| 208949 | HUHe variant click editor (CE1) - pCMV-T7-RepBM-nCas9-EcKlenow (JO606) | A variant CE1 construct with RepBM HUH endonuclease, expressed from CMV or T7 promoters. |
| 208950 | HUHe variant click editor (CE1) - pCMV-T7-FBYNV-nCas9-EcKlenow (JO610) | A variant CE1 construct with FBYNV HUH endonuclease, expressed from CMV or T7 promoters. |
| 208951 | HUHe variant click editor (CE1) - pCMV-T7-TGMV-nCas9-EcKlenow (JO659) | A variant CE1 construct with TGMV HUH enconuclease, expressed from CMV or T7 promoters. |
| 208952 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-TaqStoffel (JO595) | A variant CE1 construct with Taq Stoffel DNA polymerase, expressed from CMV or T7 promoters. |
| 208953 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-PolBeta (JO574) | A variant CE1 construct with human DNA polymerase beta, expressed from CMV or T7 promoters. |
| 208954 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-Phi29 (JO582) | A variant CE1 construct with Phi29 DNA polymerase (-exo), expressed from CMV or T7 promoters. |
| 208955 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-T7Sequenase (JO584) | A variant CE1 construct with T7 Sequenase DNA polymerase, expressed from CMV or T7 promoters. |
| 208956 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-T4DNApol (JO638) | A variant CE1 construct with T4 DNA polymerase (-exo, activity enhanced), expressed from CMV or T7 promoters. |
| 208957 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-Phi29(+exo) (LM2938) | A variant CE1 construct with Phi29 DNA polymerase (exo active), expressed from CMV or T7 promoters. |
| 208958 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-ePhi29(+exo) (LM2990) | A variant CE1 construct with ePhi29 DNA polymerase (exo active), expressed from CMV or T7 promoters. |
| 208959 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-ePhi29 (LM2993) | A variant CE1 construct with ePhi29 DNA polymerase (-exo), expressed from CMV or T7 promoters. |
| 208960 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-B103 (LM2963) | A variant CE1 construct with B103 DNA polymerase, expressed from CMV or T7 promoters. |
| 208961 | Polymerase-unfused click editor - pCMV-T7-PCV2-nCas9 (LM2702) | Polymerase-unfused click editor comprised of PCV2 fused to nCas9, expressed from CMV or T7 promoters. |
| 208962 | Polymerase-recruited click editor - pCMV-T7-PCV2-nCas9-CC(N6) (LM2705) | Polymerase-recruited click editor comprised of PCV2 fused to nCas9 with a C-terminal N6 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208963 | Polymerase expression - pCMV-T7-EcKlenow (LM2692) | Unfused EcKlenow DNA polymerase (-exo), expressed from CMV or T7 promoters. |
| 208964 | Polymerase expression - pCMV-Phi29(+exo) (LM2759) | Unfused Phi29 DNA polymerase (+exo), expressed from CMV or T7 promoters. |
| 208965 | Polymerase expression - pCMV-Phi29 (LM2708) | Unfused Phi29 DNA polymerase (-exo), expressed from CMV or T7 promoters. |
| 208966 | Polymerase expression - pCMV-ePhi29(+exo) (LM2995) | Unfused ePhi29 DNA polymerase (+exo), expressed from CMV or T7 promoters. |
| 208967 | Polymerase expression - pCMV-ePhi29 (LM2802) | Unfused ePhi29 DNA polymerase (-exo), expressed from CMV or T7 promoters. |
| 208968 | Polymerase expression - pCMV-B103 (LM2907) | Unfused B103 DNA polymerase, expressed from CMV or T7 promoters. |
| 208969 | Recruited polymerase expression - pCMV-T7-EcKlenow-CC(N5) (LM2696) | Recruited EcKlenow DNA polymerase (-exo) with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208970 | Recruited polymerase expression - pCMV-T7-Phi29(+exo)-CC(N5) (LM2761) | Recruited Phi29(+exo) DNA polymerase with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208971 | Recruited polymerase expression - pCMV-T7-Phi29-CC(N5) (LM2726) | Recruited Phi29 DNA polymerase (-exo) with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208972 | Recruited polymerase expression - pCMV-T7-ePhi29(+exo)-CC(N5) (LM2982) | Recruited ePhi29(+exo) DNA polymerase with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208973 | Recruited polymerase expression - pCMV-T7-ePhi29-CC(N5) (LM2719) | Recruited ePhi29 DNA polymerase (-exo), with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208974 | Recruited polymerase expression - pCMV-T7-B103-CC(N5) (LM2946) | Recruited B103 DNA polymerase with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters. |
| 208976 | Nickase SpCas9 - pCMV-T7-nSpCas9(H840A)-P2A-GFP (HES140) | Nickase SpCas9 (H840A) with a P2A GFP and 3xFLAG, expressed from a CMV or T7 promoters. |
| 208977 | pegRNA entry vector - pUC19-U6-[BsmBI_entry]-term (MNW320) | Entry vector for human U6 promoter driven SpCas9-based pegRNAs, comprised of hU6-[BsmBI]-terminator (spacer and RTT/PBS oligos must be cloned in) |
| 217785 | HUHe variant click editor (CE1) - pCMV-T7-BDFV-nCas9-EcKlenow (JO1271) | Variant CE1 construct with BDFV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217786 | HUHe variant click editor (CE1) - pCMV-T7-CoCV-nCas9-EcKlenow (JO1262) | Variant CE1 construct with CoCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217787 | HUHe variant click editor (CE1) - pCMV-T7-FiCV-nCas9-EcKlenow (JO1263) | Variant CE1 construct with FiCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217788 | HUHe variant click editor (CE1) - pCMV-T7-GuCV-nCas9-EcKlenow (JO1279) | Variant CE1 construct with GuCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217789 | HUHe variant click editor (CE1) - pCMV-T7-PCV1-nCas9-EcKlenow (JO1315) | Variant CE1 construct with PCV1 HUH endonuclease, expressed from CMV or T7 promoters. |
| 217790 | HUHe variant click editor (CE1) - pCMV-T7-StCV-nCas9-EcKlenow (JO1269) | Variant CE1 construct with StCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217791 | HUHe variant click editor (CE1) - pCMV-T7-CaCV-nCas9-EcKlenow (JO1261) | Variant CE1 construct with CaCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217792 | HUHe variant click editor (CE1) - pCMV-T7-Chimp17-nCas9-EcKlenow (JO1275) | Variant CE1 construct with Chim HUH endonuclease, expressed from CMV or T7 promoters. |
| 217793 | HUHe variant click editor (CE1) - pCMV-T7-GoCV-nCas9-EcKlenow (JO1277) | Variant CE1 construct with GoCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217794 | HUHe variant click editor (CE1) - pCMV-T7-SwCV-nCas9-EcKlenow (JO1270) | Variant CE1 construct with SwCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217795 | HUHe variant click editor (CE1) - pCMV-T7-RaCV-nCas9-EcKlenow (JO1268) | Variant CE1 construct with RaCV HUH endonuclease, expressed from CMV or T7 promoters. |
| 217796 | Click editor utilizing Cdc13 for template recruitment - pCMV-T7-Cdc13-nCas9-EcKlenow (JO1293) | Variant CE1 construct with Cdc13 telomere binding protein for template recruitment, expressed from CMV or T7 promoters. |
| 217797 | Click editor utilizing Cdc13(Y556A) for template recruitment - pCMV-T7-Cdc13(Y556A)-nCas9-EcKlenow (JO1329) | Variant CE1 construct with Cdc13(Y556A) telomere binding protein for template recruitment, expressed from CMV or T7 promoters. |
| 217798 | Lentiviral click editor (CE1) plasmid - LTR-pEF1a-PCV2-nCas9-EcKlenow (DRR921) | Lentiviral plasmid to produce the prototypical CE1 construct (PCV2-nCas9-EcKlenow), expressed from the EF1a promoter |
| 217799 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-M-MLV(WT) (DRR1012) | Variant CE1 construct with wild-type M-MLV revese transcriptase (RT), expressed from CMV or T7 promoters |
| 217800 | Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-M-MLV(PE2) (JO586) | Variant CE1 construct with engineered M-MLV reverse transcriptase (RT; found in PE2), expressed from CMV or T7 promoters |
| 217801 | SaCas9 variant click editor (CE1) - pCMV-T7-PCV2-nSaCas9-EcKlenow (MLE194) | Variant CE1 construct with SaCas9, expressed from CMV or T7 promoters. |
| 217802 | Click editor utilizing mSA for template recruitment - pCMV-T7-mSA-nCas9-EcKlenow (JO1391) | Variant CE1 construct with mSA for template recruitment, expressed from CMV or T7 promoters. |
| 217803 | Click editor utilizing mSA for template recruitment and ePhi29 DNA polymerase - pCMV-T7-mSA-nCas9-ePhi29 (JO1398) | Variant CE1 construct with mSA for template recruitment and ePhi29(D169A) DNA pol, expressed from CMV or T7 promoters. |
| 217804 | Unfused click editor construct; PCV2-EcKlenow fusion - pCMV-T7-PCV2-EcKlenow (JO1421) | Unfused click editor construct expressing PCV2-EcKlenow, expressed from CMV or T7 promoters. |
| 217805 | Unfused click editor construct; PCV2-ePhi29 fusion - pCMV-T7-PCV2-ePhi29 (JO1420) | Unfused click editor construct expressing PCV2-ePhi29(D169A), expressed from CMV or T7 promoters. |
| 217806 | Unfused click editor construct; mSA-EcKlenow fusion - pCMV-T7-mSA-EcKlenow (JO1401) | Unfused click editor construct expressing mSA-EcKlenow, expressed from CMV or T7 promoters. |
| 217807 | Unfused click editor construct; mSA-ePhi29 - pCMV-T7-mSA-Phi29 (JO1411) | Unfused click editor construct expressing mSA-ePhi29(D169A), expressed from CMV or T7 promoters. |
| 217808 | Click editor (CE1) mRNA expression vector - T7-5'UTR-PCV2-nCas9-EcKlenow-3'UTR-polyA (CJT472) | Expression vector for mRNA production of the click editor construct (CE1; PCV2-nCas9-EcKlenow) |