| Multi-well plate screening |
Simple |
Time consuming |
SFM4-3 |
77, 102, 104, 106 and 115
|
| Direct identification of single active mutants |
Limited screening throughput |
SFM4-6 |
|
|
SFM4-9 |
|
|
Tgo RT-TKK |
|
|
Tgo RT-C8 |
|
|
Tgo Pol6G12 |
|
|
Tgo PolC7 |
|
|
Tgo PolD4K |
|
|
Tgo RT521K |
|
|
Tgo RT521 |
|
|
KF I709E E710G |
|
|
Taq AA40 |
| CSR |
High throughput |
Target polymerase needs to replicate the full-length of its own gene |
Taq T8 |
78, 80, 81, 95, 116 and 167
|
| Simple |
High temperature is usually needed to break the emulsified cells |
Taq H15 |
|
|
Tth SΔTthCs12RsEx pol mutants |
|
|
Phi29 DNAP Mut |
|
|
Bst v5.9 |
|
|
Bst v7.16 |
|
|
KOD RTX |
|
|
KOD RTX-Ome v6 |
| spCSR |
High throughput |
High temperature is usually needed to break the emulsified cells |
Pfu DNAP E10 |
94 and 115
|
| Target polymerase only needs to replicate a part of its own gene |
|
Taq AA40 |
| Reduced adaptive burden |
|
|
| Tunable selection stringency |
|
|
| Improved selection sensitivity and versatility |
|
|
| CST |
High throughput |
High temperature is usually needed to break the emulsified cells |
Tgo Pol6G12 |
104
|
| Allows the selection for activities towards difficult nucleoside triphosphate substrates and under challenging conditions |
Plasmid DNA has to be used as the extension template for the tagging primer |
Tgo PolC7 |
|
|
Tgo PolD4K |
|
|
Tgo RT521K |
|
|
Tgo RT521 |
| CPR |
High throughput |
High temperature is usually needed to break the emulsified cells |
T7 RNAP CGG-R7-8 |
117
|
| Expanded scope of proteins to be evolved |
Challenging to design genetic circuits |
T7 RNAP CGG-R12-KIRV |
| Mitigated effect on host fitness |
|
|
| CBL |
High throughput |
High temperature is usually needed to break the emulsified cells |
Tgo RT-TKK |
102
|
| Suitable for evolving various reverse transcription activities |
Experiment complexity |
Tgo RT-C8 |
| Phage display |
High throughput |
The target polymerase needs to be actively displayed on phage |
SFM4-3 |
77, 125 and 129
|
| Kinds of the nucleic acid template, primer and nucleoside triphosphates for selection can all be well controlled |
|
SFM4-6 |
| Adjustable selection stringency |
|
SFM4-9 |
| Rapid reproduction of phage |
|
SFR1 |
|
|
SFR2 |
|
|
SFR3 |
|
|
Phi29 DNAP |
| PACE |
High throughput |
Experiment complexity |
T7 RNAP A6-36.4 |
131
|
| Rapid reproduction of phage |
Expensive facilities |
|
| Continuous evolution |
Challenging to design genetic circuits |
|
| Minimal researcher intervention |
|
|
| Rapid evolutionary cycle |
|
|
| Cell surface display |
High throughput |
The target polymerase needs to be actively displayed on cell surface |
KF I709E E710G |
106
|
| Expanded scope of polymerases to be displayed for selection |
