Table 1 Overview of microbial CBEs
Microbes | Cas domain | PAM | Editing window(nt) | Sequence preferences | High fidelity modification | Off-target testing |
|---|---|---|---|---|---|---|
(This work) E. coli S. oneidensis P. aeruginosa | HF-ID824-evoCDA-FrCas9n-UGI | 5’-NNTA | 1 ~ 19 1 ~ 18 1 ~ 18 | None | R16A/Q964A | Predicting off-target tests; orthogonal R-loop assays; whole-genome sequencing |
E. coli6 | dSpCas9-CDA-UGI-LAV | 5’-NGG | 1 ~ 5 | NA | NA | whole-genome sequencing |
E. coli12 | rAPOBEC1-SpCas9n-UGI-NLS | 5’-NGG | 3 ~ 8 | NA | NA | NA |
E. coli16 | rAPOBEC1-dLbCas12a-UGI | 3’-TTTV | 7 ~ 13 | NA | NA | Predicting off-target sites |
E. coli62 | GCN4-nSpCas9-scFV-APOBEC-UGI | 5’-NGG | 1 ~ 14 | NA | NA | Predicting off-target sites |
S. oneidensis13 | APOBEC1-nSpCas9 | 5’-NGG | 3 ~ 8 | NA | NA | NA |
S. oneidensis23 | nSpCas9-AID | 5’-NGG | 1 ~ 6 | TC > AC > CC > GC | NA | NA |
P. aeruginosa7 | APOBEC1-nCas9-UGI | 5’-NGG | 2 ~ 9 | TC > AC > CC > GC | NA | whole-genome sequencing |
P. aeruginosa68 | APOBEC1-SPRYnCas9-UGI-UGI | 5’-NRN | 3 ~ 8 | NA | NA | whole-genome sequencing |
C. glutamicum69 | nVQR-SpCas9-AID nVRER-SpCas9-AID | 5’-NGAN 5’-NGCG | 1 ~ 5 1 ~ 5 | NA | NA | Predicting off-target sites |
B. subtilis70 | CDA-nSpCas9 | 5’-NGG | 1 ~ 5 | NA | NA | whole-genome sequencing |
B. subtilis8 | CDA-nSpCas9 | 5’-NGG | −2 ~ 6 | NA | NA | NA |
B. subtilis63 | CDA-dBhCas12b-UGI-UGI | 3’-TTN | 1 ~ 19 | NA | NA | Predicting off-target sites |
A. hydrophila20 | APOBEC-nSpCas9 | 5’-NGG | 3 ~ 8 | TC > AC or CC > GC | NA | NA |
K. pneumoniae21 | APOBEC-nSpCas9 | 5’-NGG | 3 ~ 8 | TC > CC/AC > GC | NA | Predicting off-target sites |
G. sulfurreducens19 | APOEBC-nSpCas9-UGI | 5’-NGG | 3 ~ 8 | TC > CC/AC > GC | NA | NA |
