Table 1.
Mutation efficiency of the type II CRISPR system in P. putida KT2440
| Host cell | gRNA plasmid | Deletion | Replacement | Insertion | Size | Results | Plasmid curing efficiency |
|---|---|---|---|---|---|---|---|
| KT2440 harboring pCAS-RK2K | pSEVA-gRic6T | nicC (PP _3944) | 1149 | 15/15 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gRic5T | nicC (PP _3944) | 1149 | 39/45 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gRic6TΔNicC::rhla | nicC (PP _3944) | rhla | 888 | 38/40 | ND | |
| KT2440 harboring pCAS-RK2K | pSEVA-gRic6TΔNicC::dCas9 | nicC (PP _3944) | dCas9 | 4107 | 20/20 | ND | |
| KT2440 harboring pCAS-RK2K | pSEVA-gRic6TΔNicC::T7 RNA polymerase | nicC (PP _3944) | T7 | 4518 | 14/20 | 5/5 | |
| KT2440 harboring pCAS-RK2K | pSEVA-gRic6PAM2 | Single nucleotide | Single nucleotide | 1 | 5/5 | ND | |
| KT2440 harboring pCAS-RK2K | pSEVA-NicA20 | Single nucleotide | Single nucleotide | A20 | 21 | 7/7 | ND |
| KT2440 harboring pCAS-RK2K | pSEVA-NicA21 | A20 | 20 | 5/5 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gR0552T | PP_0552 | 1089 | 8/10 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gR3361T | PP_3361 | 3048 | 11/13 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gR3733T | PP_3733 | 1059 | 18/18 | ND | ||
| KT2440 harboring pCAS-RK2K | pSEVA-gR3889T | PP_3889 | 1299 | 11/12 | ND | ||
| KT2440 harboring pCAS-RK2T | pSEVA-gR3947-3948T | PP_3947–PP_3948 | 4033 | 18/18 | 5/5 | ||
| KT2440 harboring pCAS-RK2T | pSEVA-gR3939-3940T | PP_3939–PP_3940 | 2743 | 9/10 | 5/5 | ||
| KT2440 harboring pCAS-RK2T | pSEVA-gR3846T | PP_3846 | 816 | 10/10 | ND | ||
| KT2440 harboring pCAS-RK2T | pSEVA-gR1706T | PP_1706 | 363 | 15/16 | ND | ||
| KT2440 harboring pCAS-RK2T | pSEVA-gR5301T | PP_5301 | 264 | 10/10 | ND | ||
| KT2440 harboring pCpf1-RK2K | pSEVA-gcR3361T | PP_3361 | 3048 | 4/4 | ND | ||
| KT2440 harboring pCpf1-RK2K | pSEVA-gcR5301T | PP_5301 | 264 | 9/9 | 2/2 |
rhla was cloned from Pseudomonas aeruginosa PAOl, and T7 RNA polymerase was amplified from Escherichia coli BL21(DE3)
ND not determined