Table 1.
Summary of phage mutants engineered by CRISPR-Cas13a.
| Phage | Gene No./Genomic site | Identified protein | Modification | % plaques acrVIA1+1 (n2) | Desired mutant isolated? |
|---|---|---|---|---|---|
| ФKZ | Upstream of orf54 | - | Insertion | 41.2% (17) | Yes |
| Downstream of orf120 | - | Insertion | 50.0% (16) | ||
| orf39 | PhuZ | Deletion | 52.9% (17) | ||
| mNeonGreen-PhuZ fusion | 25.0% (16) | ||||
| mCherry-PhuZ fusion |
25.0% (16) | ||||
| orf93 | Inner body protein | Deletion | 20.0% (20) | ||
| gp93-mNeonGreen fusion | 18.2% (22) | ||||
| orf241 | Hypothetical | Deletion | 26.1% (23) | ||
| orf241, orf242 | Hypothetical | Double deletions | 41.7% (12) | ||
| orf54 | Shell | Deletion | 20.0% (15) | No | |
| mCherry-gp54 fusion |
8.3% (24) | ||||
| gp54-mCherry fusion |
9.1% (11) | ||||
| gfp11-gp54 fusion | 7.0% (57) | ||||
|
orf89, orf90,
orf91, orf92, orf93 |
Inner body proteins | Deletion | 11.8% (17) | ||
| orf146 | Structural protein | Deletion | 0% (89) | ||
| OMKO1 | Downstream of the capsid gene | - | Insertion with a barcode | 70.8% (24) | Yes |
| Upstream of the shell gene | - | Insertion | 50.0% (12) | ||
| PaMx41 | orf24 | Hypothetical | Deletion | 100.0% (8) | Yes |
1
Percentage of PCR-identified recombinants out of all tested plaques.
2
Number of plaques analyzed by PCR to screen for recombinants.