Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.
da Silva JF, Tou CJ, King EM, Eller ML, Ma L, Rufino-Ramos D, Kleinstiver BP
bioRxiv. 2023 Sep 13:2023.09.12.557440. doi: 10.1101/2023.09.12.557440. Preprint. PubMed Article
da Silva JF, Tou CJ, King EM, Eller ML, Ma L, Rufino-Ramos D, Kleinstiver BP
bioRxiv. 2023 Sep 13:2023.09.12.557440. doi: 10.1101/2023.09.12.557440. Preprint. PubMed 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) |