Table 1 Current CFES specific for phages’ production
From: Cell-free expression system: a promising platform for bacteriophage production and engineering
Year of publication | Phage bacterial host | Type of host | Cell extract preparatory host | Genetic/System modifications | Phage | Phage titer (PFU/mL) | Reference |
|---|---|---|---|---|---|---|---|
2024 | E. coli | Gram-negative | E. coli JM109 | Addition of 15µL GamS (150 mM) 100 ng gDNA | M13 | 105 | Xu et al. [66] |
2024 | E. coli | Gram-negative | E. coli JM109 cells harboring the helper phage plasmid | Addition of 15µL GamS (150 mM) a. Phagemid b. Phagemid with the pMB1 origin of replication | M13 | a. 108 a. 109 | Xu et al. [66] |
2024 | E. coli B | Gram-negative | E. coli BL21-∆recBCD Rosetta2 | T7 genome is re-assembled from long PCR fragments to allow for genetic manipulations in the study. “GenBank consecutive accession codes PP384393 to PP384410”. | T7 (Boca Scientific, #310025) | 1010-1011 | Levrier et al. [57] |
2024 | E. coli B | Gram-negative | E. coli BL21-∆recBCD Rosetta2 | Phage T6 169 kbp GenBank: NC_054907 | ~ 108 | Levrier et al. [57] | |
2024 | E. coli BE [67] | Gram-negative | E. coli BL21-∆recBCD Rosetta2 | Phage VpaE1 88 kbp GenBank: NC_027337.1 | ~ 1010 | Levrier et al. [57] | |
2024 | Salmonella LT2 | Gram-negative | E. coli BL21-∆recBCD Rosetta2 | Phage FelixO1 86 kbp GenBank: NC_005282 | ~ 108 | Levrier et al. [57] | |
2024 | Salmonella LT2 | Gram-negative | E. coli BL21-∆recBCD Rosetta2 | Phage S16 160 kbp GenBank: NC_020416 | ~ 108 | Levrier et al. [57] | |
2024 | Salmonella enterica subsp. enterica serovar Typhimurium strain LT2 | Gram-negative | E. coli | a. Without supplemental energy mix b. With an energy mix which includes dNTPs | Phage FelixO1 86 kbp GenBank: NC_005282 | a. 5.4 × 107 b. ~4.0 × 108 | McFarlane et al. [68] |
2024 | E. coli strain BE | Gram-negative | E. coli | Without supplemental energy mix | VpaE1 | 7.5 × 1010 | McFarlaneet al. [68] |
2023 | E. coli BL21 ATCC BAA-1025 | Gram-negative | E. coli BL21 ATCC BAA-1025 | Overexpression of inf C, OxyS, and CyaR | T7 (ATCC BAA-1025-B2) | ~ 108-109 | Brooks et al. [28] |
2023 | E. coli BL21 ATCC BAA-1025 | Gram-negative | E. coli BL21 ATCC BAA-1025 | Repression of recC, and rna | T7 ATCC BAA-1025-B2) | ~ 107-109 | Brooks et al. [28] |
2022 | E. coli K603, F+ | Gram-negative | E. coli BL12 (DE3) | pQβ7 plasmid or Qβ (+)-RNA 3% PEG 6000 Cell extract of 24.3 mg/mL protein | Qβ Shaklee et al. [69] | ~ 106 | Rasmussen et al. [70] |
2022 | E. coli EV36 | Gram-negative | E. coli Rosetta 2 transformed with pAD-LyseR plasmid | Using CRISPER/Cas 9 selection and SpyTag/SpyCatcher system to attach proteins to the minor capsid protein | K1F & K1F-GFP-SpyTag | 109 | Liyanagedera et al. [24] |
2022 | E. coli DSM 613 | Gram-negative | E. coli Rosetta 2 (DE3) | Addition of 2µL GamS (150 µM) | T7 phage, DSM 4623 | 1012-1011 | Emslander et al. [52] |
2022 | E. coli DSM 613 | Gram-negative | E. coli Rosetta 2 (DE3) | Addition of 2µL GamS (150 µM) | MS2 phage, DSM 13767 | 1012-1011 | Emslander et al. [52] |
2022 | Bacillus subtilis DSM 5547 | Gram-positive | E. coli Rosetta 2 (DE3) | B. subtilis housekeeping sigma factor SigA Addition of 2µL GamS (150 µM) | Phi29 (NCBI ID: NC_011048.1) DSM 5546 | 1010 | Emslander et al. [52] |
2022 | Bacillus subtilis DSM 402 | Gram-positive | E. coli Rosetta 2 (DE3) | B. subtilis housekeeping sigma factor SigA Addition of 2µL GamS (150 µM) | Goe1 (vB_BsuP_- Goe1, NCBI ID NC_049975.1) DMS 101030 | 1010 | Emslander et al. [52] |
2022 | Yersinia pestis EV76 | Gram-negative | E. coli Rosetta 2 (DE3) | Addition of 2µL GamS (150 µM) | PhiA1122 (NCBI ID: NC_004777.1) [71] | 108 | Emslander et al. [52] |
2022 | Enteroaggregative E. coli [72] | Gram-negative | E. coli Rosetta 2 (DE3) | Addition of 2µL GamS (150 µM) | CLB-P3 (GeneBank: OL800706) | 1010 | Emslander et al. [52] |
2022 | Klebsiella pneumoniae (Bw1) | Gram-negative | E. coli Rosetta 2 (DE3) | Addition of 2µL GamS (150 µM) | vB_KpS_Muc5 (GeneBank: OM687892) | 108 | Emslander et al. [52] |
2021 | E. coli | Gram-negative | E. coli Rosetta 2 (DE3) | Using 5 sonication cycles and 0.5 mg/mL lysozyme | T7 | Up to 109 | Falgenhauer et al. [53] |
2021 | E. coli B | Gram-negative | E. coli BL21 Rosetta 2 | Addition of 3 µM of chi6 short DNA Addition of 0.1 mM dNTPs PEG8000, 3.5% (4.3 mM) | T7 (Boca Scientific) | 1013 | Garenne et al. [44] |
2018 | E. coli B | Gram-negative | E. coli BL21 Rosetta 2 | 1 nM gDNA Magnesium glutamate 4–7 mM Potassium glutamate 40–80 mM PEG800, 3% v/v | T4 | 108 | Rustad et al. [30] |
2017 | E. coli B | Gram-negative | E. coli BL21 Rosetta 2 | Addition of 0.5 µM Chi6 short DNA 0.25 nM gDNA | T7 (Boca Scientific) | ~ 1011 | Marshall et al. [73] |
2016 | E. coli B | Gram-negative | E. coli | Addition of dNTPs 0.25 nM gDNA | T7 (NCBI ID: NC_001604.1) | 3.35 × 1011 | Garamella et al. [19] |
2016 | E. coli HF4714 | Gram-negative | E. coli | 5 nM gDNA | PhiX174 (GenBank: J02482.1) | 1.9 × 1012 | Garamella et al. [19] |
2016 | E. coli C-1 | Gram-negative | E. coli | 150 nM gDNA | MS2 (NCBI ID: NC_001417.2) | 4.23 × 1012 | Garamella et al. [19] |
2012 | E. coli C | Gram-negative | E. coli BL21 Rosetta 2 | Addition of 0.5 nM dNTPS 30 nM gDNA 3.3 µM GamS | PhiX174 (New England Biolabs) | ~ 106 | Shin et al. [74] |
2012 | E. coli B | Gram-negative | E. coli BL21 Rosetta 2 | Addition of 0.5 nM dNTPS 1 nM gDNA 3.3 µM GamS | T7 (Boca Scientific) | ~ 1010 | Shin et al. [74] |