John van der Oost Lab: Size and sequence variation of the Lid of the Cas12a nuclease domain results in a nickase phenotype
Unpublished
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 262848 | pCas-LbCas12a-K932G-N933G-S934A-R935G_pCB482-p15A | LbCas12a Lid variant K932G/N933G/S934A/R935G, constitutively expressed from PJ23108 in E. coli. Used in the in vivo nicking screen. Chloramphenicol, p15A. |
| 262849 | pCas-LbCas12a-Q888P-N889P_pCB482-p15A | LbCas12a bridge-helix double-proline variant Q888P/N889P, adapted from FnoCas12a KD2P (Parameshwaran 2021 FEBS Lett). Constitutive PJ23108. Chloramphenicol, p15A. |
| 262850 | pCas-LbCas12a-Lid2_pCB482-p15A | LbCas12a Lid2 nickase (G930Y/F931C/K932S/N933S plus deletion of 934-941), constitutively expressed from PJ23108 in E. coli. Nicks the non-target strand. Chloramphenicol, p15A. |
| 262851 | pET-6xHis-LbCas12a-Lid2 | N-terminally His6-tagged LbCas12a Lid2 nickase for IPTG-inducible expression and purification from E. coli Rosetta (DE3). Kanamycin, T7/lacO. |
| 262852 | pGFP-PS1-sgRNA1-reporter-SpCas9_pSC101-RepA-N99D | GFP reporter carrying protospacer PS1 and a constitutive SpCas9 sgRNA cassette, for the in vivo nicking screen in E. coli. Kanamycin, pSC101 RepA N99D. |
| 262853 | pGFP-PS1-crRNA1-reporter-LbCas12a_pSC101-RepA-N99D | GFP reporter carrying protospacer PS1 and a constitutive LbCas12a crRNA array, for the in vivo nicking screen in E. coli. Kanamycin, pSC101 RepA N99D. |
| 262854 | pRFP-PS2-sgRNA2-reporter-SpCas9_pBR322 | RFP reporter carrying protospacers PS1 and PS2 and a constitutive SpCas9 sgRNA cassette, for the in vivo nicking screen in E. coli. Ampicillin, pBR322. |
| 262855 | pRFP-PS2-crRNA2-reporter-LbCas12a_pBR322 | RFP reporter carrying protospacers PS1 and PS2 and a constitutive LbCas12a crRNA array, for the in vivo nicking screen in E. coli. Ampicillin, pBR322. |