Journal: bioRxiv

Article Title: CRISPR-Cas12a targeting of ssDNA plays no detectable role in immunity

doi: 10.1101/2022.03.10.483831

Figure Lengend Snippet: Cas12a targeting does not elicit abortive infection or prevent the rise of phage esapers. (A) Growth curves of P. aeruginosa PAO1 tn7::LbCas12a strain expressing targeting or non-targeting crRNAs infected with ϕDMS3 at multiplicities of infection (MOI) indicated in legend. (B) Phage plaque assays with ten-fold serial dilutions of wildtype or mutated ϕ JBD30 phage. Bacterial clearance (black) indicates phage replication. Sequences of the targeted phage gene show mutated nucleotides (magenta) in phages able to escape Cas12a targeting. PAM, protospacer adjacent motif (teal); WT, wildtype phage; ESC, escaper phage.

Article Snippet: To generate the PAO1 tn7::LbCas12a strain (Supplementary Figure S1), LbCas12a was first amplified from pMBP-LbCas12a (Addgene #113431) and cloned into pUC18-mini-Tn7T-lac. pUC18-Tn7T-LbCas12a was co-electroporated with pTNS3 plasmid into the P. aeruginosa PAO1 strain according to previously published protocols ( , ).

Techniques: Infection, Expressing

Journal: bioRxiv

Article Title: CRISPR-Cas12a targeting of ssDNA plays no detectable role in immunity

doi: 10.1101/2022.03.10.483831

Figure Lengend Snippet: Cas12a does not detectably cis-cleave invasive ssDNA in bacteria. (A) Conjugation efficiency for plasmid cis-targeting in P. aeruginosa PAO1 strain. During conjugation, the donor is nicked and the leading strand (LDS) is transferred as ssDNA to the wildtype (-LbCas12a) or LbCas12a-expressing (+LbCas12a) recipient cell via the mating pilus. LbCas12a was co-expressed with crRNAs complementary to a DMS3 protospacer cloned into the leading (LDS) or lagging (LGS) strand, with (+) or without (-) the correct protospacer adjacent motif (PAM). T/D indicates the ratio of transconjugants to donors. Data are presented as mean±SEM (n = 3). LDS, complementary protospacer is on leading strand; LGS, complementary protospacer is on lagging strand; NP, no protospacer on plasmid. (B) Plaque assay for phage cis-targeting in E. coli BW25113 F’ strain. Bacterial lawns were infected with dsDNA λ vir or with M13, which enters and leaves the cell as ssDNA but replicates via a dsDNA intermediate. LbCas12a was co-expressed in bacteria with crRNA complementary to M13 positive strand (without PAM) or λ vir (with PAM) during phage infection. Bacterial clearance (black) indicates phage replication. (C) In vitro LbCas12a cleavage assay on dsDNA template encoding DMS3 protospacer or M13 ssDNA.

Article Snippet: To generate the PAO1 tn7::LbCas12a strain (Supplementary Figure S1), LbCas12a was first amplified from pMBP-LbCas12a (Addgene #113431) and cloned into pUC18-mini-Tn7T-lac. pUC18-Tn7T-LbCas12a was co-electroporated with pTNS3 plasmid into the P. aeruginosa PAO1 strain according to previously published protocols ( , ).

Techniques: Bacteria, Conjugation Assay, Plasmid Preparation, Expressing, Clone Assay, Plaque Assay, Infection, In Vitro, Cleavage Assay

Journal: bioRxiv

Article Title: CRISPR-Cas12a targeting of ssDNA plays no detectable role in immunity

doi: 10.1101/2022.03.10.483831

Figure Lengend Snippet: Cas12a does not trans-cleave invasive ssDNA in bacteria. (A) Right: Schematic for plasmid coconjugation and cleavage in P. aeruginosa PAO1 strain. Two plasmids were conjugated simultaneously from different donors into the same recipient cell. LbCas12a was co-expressed in the recipient cell with crRNA complementary to pTarget but not pTrans. A parallel co-conjugation mating using pTrans and pTarget(-PS) (identical to pTarget but lacking the protospacer and PAM) was carried out as a control. Conjugation efficiency was assessed using antibiotic selection specific to each plasmid and represented as the transconjugant to donor ratio (T/D). Data are presented as mean ± SEM ( n = 3); ** p =0.0036 by unpaired, two-sided Student’s t test; ns, not significant ( p >0.05) (B) Right: Schematic for phage co-infection in E. coli. Bacterial lawns of E. coli BW25113 F’ strain co-expressing LbCas12a and λ-targeting or non-targeting crRNA were singly infected or co-infected with ssDNA phage M13 and dsDNA phage λvir. Strains were infected with either low titer (low), high titer (high), or ten-fold serial dilutions (serial) of phage as indicated.

Article Snippet: To generate the PAO1 tn7::LbCas12a strain (Supplementary Figure S1), LbCas12a was first amplified from pMBP-LbCas12a (Addgene #113431) and cloned into pUC18-mini-Tn7T-lac. pUC18-Tn7T-LbCas12a was co-electroporated with pTNS3 plasmid into the P. aeruginosa PAO1 strain according to previously published protocols ( , ).

Techniques: Bacteria, Plasmid Preparation, Conjugation Assay, Control, Selection, Infection, Expressing

Journal: medRxiv

Article Title: Altered neutrophil G-protein receptor signalling linked to impaired chemotaxis and increased ROS and NET production in older people with frailty

doi: 10.64898/2025.12.16.25342352

Figure Lengend Snippet: (A) ROS production in response to fMLP in neutrophils primed with GM-CSF (5ng/mL), TNFa (10ng/mL) or unprimed (UP). (B) NET production in response to PMA (0.1mg/mL), A23187 (3.8mM ), LPS (1 or 10 ng/mL) or unstimulated (cells only). Purple = FR (n=5-10), Pink = RA (n=5-9), Orange = HO (n=5-10), Green = HY (n=5-7). Analysis by ANOVA (*p<0.05, **p<0.01, ***p>0.001). (C) Representative microscopy images of NETs on coverslips (Blue = DNA, Red = MPO, Green = neutrophil elastase). Scale bar 50μM.

Article Snippet: Cells were allowed to settle for 15min prior to addition of LPS from P. aeruginosa (1 or 10ng/mL, Merck), PMA (0.1mg/mL, Merck), or A23187 (3.8μM, Merck) along with an untreated control.

Techniques: Microscopy

Journal: medRxiv

Article Title: Altered neutrophil G-protein receptor signalling linked to impaired chemotaxis and increased ROS and NET production in older people with frailty

doi: 10.64898/2025.12.16.25342352

Figure Lengend Snippet: Neutrophils were either untreated (UT) or primed with TNFα (10ng/mL) before incubating with S. aureus bacteria (10:1 ratio) for up to 2h. Purple = FR (n=5-10), Pink = RA (n=5-9), Orange = HO (n=5-10), Green = HY (n=5-7). Analysis by ANOVA (*p<0.05, **p<0.01, ***p>0.001).

Article Snippet: Cells were allowed to settle for 15min prior to addition of LPS from P. aeruginosa (1 or 10ng/mL, Merck), PMA (0.1mg/mL, Merck), or A23187 (3.8μM, Merck) along with an untreated control.

Techniques: Bacteria

Journal:

Article Title: Protection against Pseudomonas aeruginosa Chronic Lung Infection in Mice by Genetic Immunization against Outer Membrane Protein F (OprF) of P. aeruginosa

doi: 10.1128/IAI.69.5.3510-3515.2001

Figure Lengend Snippet: Immunoblot of proteins extracted from whole cells of the PAO1 strain of P. aeruginosa with polyclonal anti-OprF sera obtained from DNA-immunized (pVR1020/oprF) mice. Note that the antisera reacted at bands corresponding to F (OprF), F′ (denatured OprF), and H (OprH).

Article Snippet: The following strains of P. aeruginosa were used: Fisher-Devlin (FD) immunotype 1 (ATCC 27584; Difco 0–6), FD immunotype 2 (ATCC 27313; Difco 0–11), FD immunotype 3 (ATCC 27314; Difco 0–2), FD immunotype 4 (ATCC 27315; Difco 0–1), FD immunotype 5 (ATCC 27585; Difco 0–10), FD immunotype 6 (ATCC 27579; Difco 0–7 and 0–8), FD immunotype 7 (PAO1; Difco 0–5), and KG1077, which is a protein F-deficient mutant of PAO1.

Techniques: Western Blot

Journal:

Article Title: Protection against Pseudomonas aeruginosa Chronic Lung Infection in Mice by Genetic Immunization against Outer Membrane Protein F (OprF) of P. aeruginosa

doi: 10.1128/IAI.69.5.3510-3515.2001

Figure Lengend Snippet: Quantitation of phagocytic uptake by PMNs of P. aeruginosa FD 4 cells exposed to antisera from immunized mice

Article Snippet: The following strains of P. aeruginosa were used: Fisher-Devlin (FD) immunotype 1 (ATCC 27584; Difco 0–6), FD immunotype 2 (ATCC 27313; Difco 0–11), FD immunotype 3 (ATCC 27314; Difco 0–2), FD immunotype 4 (ATCC 27315; Difco 0–1), FD immunotype 5 (ATCC 27585; Difco 0–10), FD immunotype 6 (ATCC 27579; Difco 0–7 and 0–8), FD immunotype 7 (PAO1; Difco 0–5), and KG1077, which is a protein F-deficient mutant of PAO1.

Techniques: Quantitation Assay, Bacteria, Plasmid Preparation, Control

Journal:

Article Title: Protection against Pseudomonas aeruginosa Chronic Lung Infection in Mice by Genetic Immunization against Outer Membrane Protein F (OprF) of P. aeruginosa

doi: 10.1128/IAI.69.5.3510-3515.2001

Figure Lengend Snippet: Scoring of macroscopic lung lesions in immunized mice following challenge with FD 4 P. aeruginosa in a chronic pulmonary infection model

Article Snippet: The following strains of P. aeruginosa were used: Fisher-Devlin (FD) immunotype 1 (ATCC 27584; Difco 0–6), FD immunotype 2 (ATCC 27313; Difco 0–11), FD immunotype 3 (ATCC 27314; Difco 0–2), FD immunotype 4 (ATCC 27315; Difco 0–1), FD immunotype 5 (ATCC 27585; Difco 0–10), FD immunotype 6 (ATCC 27579; Difco 0–7 and 0–8), FD immunotype 7 (PAO1; Difco 0–5), and KG1077, which is a protein F-deficient mutant of PAO1.

Techniques: Infection, Mouse Assay, Plasmid Preparation, Control

Journal:

Article Title: Protection against Pseudomonas aeruginosa Chronic Lung Infection in Mice by Genetic Immunization against Outer Membrane Protein F (OprF) of P. aeruginosa

doi: 10.1128/IAI.69.5.3510-3515.2001

Figure Lengend Snippet: Quantitation of P. aeruginosa present in the lungs of immunized mice following challenge with FD 4 P. aeruginosa in a chronic pulmonary infection model

Article Snippet: The following strains of P. aeruginosa were used: Fisher-Devlin (FD) immunotype 1 (ATCC 27584; Difco 0–6), FD immunotype 2 (ATCC 27313; Difco 0–11), FD immunotype 3 (ATCC 27314; Difco 0–2), FD immunotype 4 (ATCC 27315; Difco 0–1), FD immunotype 5 (ATCC 27585; Difco 0–10), FD immunotype 6 (ATCC 27579; Difco 0–7 and 0–8), FD immunotype 7 (PAO1; Difco 0–5), and KG1077, which is a protein F-deficient mutant of PAO1.

Techniques: Quantitation Assay, Infection, Mouse Assay, Plasmid Preparation, Control

Journal: BMC Pulmonary Medicine

Article Title: Serum (1→3)-β-D-glucan and galactomannan levels in patients with cystic fibrosis: a retrospective cohort study

doi: 10.1186/s12890-018-0614-8

Figure Lengend Snippet: Patient demographics, clinical and microbiological results stratified after serum BDG and GM positivity

Article Snippet: Serum samples were tested for Aspergillus -specific antibodies (Aspergillose Fumouze, Fumouze Diagnostics, France), Aspergillus -specific IgE, recombinant Aspergillus antigens f4 and f6 [rAsp f4 IgE, rAsp f6 IgE] (ImmunoCAP 250, Thermo Fisher Scientific, Sweden) and IgG against alkaline protease, elastase and exotoxin A of P. aeruginosa (Mediagnost GmbH, Germany).

Techniques: Sampling

Journal: BMC Pulmonary Medicine

Article Title: Serum (1→3)-β-D-glucan and galactomannan levels in patients with cystic fibrosis: a retrospective cohort study

doi: 10.1186/s12890-018-0614-8

Figure Lengend Snippet: Patient demographics, microbiological and clinical results stratified after FEV 1 at serum sampling and persistent A. fumigatus detection

Article Snippet: Serum samples were tested for Aspergillus -specific antibodies (Aspergillose Fumouze, Fumouze Diagnostics, France), Aspergillus -specific IgE, recombinant Aspergillus antigens f4 and f6 [rAsp f4 IgE, rAsp f6 IgE] (ImmunoCAP 250, Thermo Fisher Scientific, Sweden) and IgG against alkaline protease, elastase and exotoxin A of P. aeruginosa (Mediagnost GmbH, Germany).

Techniques: Sampling

Journal: BMC Pulmonary Medicine

Article Title: Serum (1→3)-β-D-glucan and galactomannan levels in patients with cystic fibrosis: a retrospective cohort study

doi: 10.1186/s12890-018-0614-8

Figure Lengend Snippet: Multivariate analysis with FEV1 predicted at serum sampling as dependent variable

Article Snippet: Serum samples were tested for Aspergillus -specific antibodies (Aspergillose Fumouze, Fumouze Diagnostics, France), Aspergillus -specific IgE, recombinant Aspergillus antigens f4 and f6 [rAsp f4 IgE, rAsp f6 IgE] (ImmunoCAP 250, Thermo Fisher Scientific, Sweden) and IgG against alkaline protease, elastase and exotoxin A of P. aeruginosa (Mediagnost GmbH, Germany).

Techniques: Sampling

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: Cross-reactivity analysis of 6xHis-tagged scFv by whole-cell ELISA and WB. Binding activities of yPac1A8 and yPgi3G4 scFvs were determined by whole-cell ELISA on a plate coated with the indicated bacteria (A): P. acidipropionici TISTR 442, P. freudenreichii TISTR 446, P. acnes DMST 14916, P. acnes strains DSM 1897, DSM 16379, DSM 30738, DSM 30753, DSM 30919, P. putida TISTR 1522, P. fluorescens TISTR 358, P. aeruginosa DMST 37186, P. aeruginosa strains TISTR 357, TISTR 781, TISTR 1101, TISTR 1287, and wells without immobilized bacteria. The bars represent the average OD values of triplicate samples, and error bars represent the standard error of the mean. A picture of the ELISA plate is shown in the Supporting Information ( Figure S1 ). (B) SDS-PAGE and WB of boiled antigen preparation of P. acnes strains, immunoblotted with yPac1A8 scFv: DMST 14916, DSM 1897, DSM 16379, DSM 30738, DSM 30753, and DSM 30919: Coomassie Brilliant Blue-stained SDS-PAGE gel. The P. acnes antigen is located between 20 and 25 kDa (arrow pointed) and was detected by yPac1A8 scFv. (C) SDS-PAGE and WB of boiled antigen preparation of P. aeruginosa strains, immunoblotted with yPgi3G4 scFv as indicated: DMST 37186, TISTR 357, TISTR 781, TISTR 1101, and TISTR 1287. The P. aeruginosa antigen located between 37 and 50 kDa (arrow pointed) was detected by yPgi3G4 scFv. Precision Plus All Blue Prestained Protein Standards (BioRad#1610373, U.S.A.) were used as a molecular weight marker.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Bacteria, SDS Page, Staining, Molecular Weight, Marker

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: Checkerboard titration and flow cytometry analysis against whole cell bacteria. The limit of detection of yPac1A8 for P. acnes DMST 14916 (A) and yPgi3G4 for P. aeruginosa DMST 37186 (C) was determined by checkerboard titration of whole-cell ELISA, using a dilution series of scFv and serial dilutions of bacteria as indicated. The lines represent the average absorbance values of duplicate samples, and error bars represent the standard error of the mean. Pictures of ELISA plates are shown in the Supporting Information ( Figures S2 and S3 ). Flow cytometry analysis of yPac1A8 (B) and yPgi3G4 scFv (D) binding to live P. acnes DMST 14916 and P. aeruginosa DMST 37186, respectively. Reactivity of 6xHis-tagged scFv is indicated by the red line. The secondary antibody alone (control) is indicated by the black line.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Titration, Flow Cytometry, Bacteria, Enzyme-linked Immunosorbent Assay, Binding Assay, Control

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: CLSM of yPac1A8 and planktonic bacteria. P. acnes DMST 14916, P. acidipropionici TISTR 442, P. freudenreichii TISTR 446, and P. aeruginosa DMST 37186 were incubated with yPac1A8 scFv, detected by Dylight 488-labeled anti-hexa-histidine mouse monoclonal antibody, and counterstained with DAPI. Photos were taken by an Apo TIRF 60x Oil DIC N2 objective of Nikon A1R confocal laser microscope. Scale bars represent 10 μm. In bright field panel (A), individual bacterial cells were seen. In DAPI panel (B), nucleoids of bacteria were stained blue. In Dylight 488 panel (C), only P. acnes DMST 14916 was stained green. In merged panel (D), the blue nucleoids and the green cell surfaces of P. acnes overlapped. In the case of P. acidipropionici , P. freudenreichii , and P. aeruginosa , only the blue nucleoids were stained.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Bacteria, Incubation, Labeling, Microscopy, Staining

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: CLSM of biofilms stained by indicated scFv. Biofilms of P. acnes DMST 14916, P. acidipropionici TISTR 442, P. putida TISTR 1522, and P. aeruginosa DMST 37186 were stained by yPac1A8 and yPgi3G4 scFvs as indicated. Scale bars represent 10 μm. In bright field panel (A), bacterial cell clusters were seen. In DAPI panel (B), nucleoids of bacteria were stained blue. In Dylight 488 panel (C), P. acnes were stained green by yPac1A8 scFv but not P. acidipropionici . Likewise, P. aeruginosa were stained green by yPgi3G4 scFv but not P. putida . In merged panel (D), the blue nucleoids and the green cell surfaces of P. acnes and P. aeruginosa overlapped. In the case of P. acidipropionici and P. putida , only the blue nucleoids were stained.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Staining, Bacteria

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: CLSM of yPgi3G4 and planktonic bacteria. Photos of P. aeruginosa DMST 37186, P. putida TISTR 1522, P. fluorescens TISTR 358, and P. acnes DMST 14916 were taken at the same specifications as in Figure . Scale bars represent 10 μm. In bright field panel (A), bacterial cells are seen. In DAPI panel (B), nucleoids of bacteria are blue. In Dylight 488 panel (C), only P. aeruginosa are green. In merged panel (D), the blue nucleoids and the green cell surfaces of P. aeruginosa overlapped. In the case of P. putida , P. fluorescens and P. acnes , only the blue nucleoids were stained.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Bacteria, Staining

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: TEM of P. acnes and P. aeruginosa . Four P. acnes DMST 14916 cells (A) and six P. aeruginosa DMST 37186 cells (B) were treated with yPac1A8 and yPgi3G4 scFvs, respectively. Photos were taken by the Hitachi Hi-Tech HT7700 transmission electron microscope at x25.0 k magnification with Zoom-1 lens mode at the accelerating voltage of 80.0 kV. Scale bars represent 1.0 μm. The large round structures are bacterial cells, and small dense black dots are 10 nm Ni-NTA nanogold particles.

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Transmission Assay, Microscopy

Journal: ACS Omega

Article Title: Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections

doi: 10.1021/acsomega.0c05340

Figure Lengend Snippet: Summary of Affinity Selection Results against P. acnes and P. aeruginosa

Article Snippet: Another study group used the recombinant exotoxin A of P. aeruginosa to do affinity selection against a human scFv phage display library based on 60 Thai blood donors and isolated anti-exotoxin A scFv clones., To the best of our knowledge, only the anti-Psl scFv of MedImmune Ltd. group has been developed into an anti-pseudomonal drug candidate.

Techniques: Selection, Clone Assay, Enzyme-linked Immunosorbent Assay, DNA Sequencing, Sequencing

Journal: Nature Communications

Article Title: Compensatory evolution of Pseudomonas aeruginosa ’s slow growth phenotype suggests mechanisms of adaptation in cystic fibrosis

doi: 10.1038/s41467-021-23451-y

Figure Lengend Snippet: The cartoon represents the trajectories of evolution from naïve to adapted strain occurring in the CF patients. Several cellular components such as the flagellum, the type 4 pili (T4 pili), the type 3 secretion system (T3SS) and numerous virulence factors are lost during within-patient evolution due to accumulation of mutations. The Gac/Rsm regulatory system coordinates the switch between acute and chronic phenotype through changes of gene expression profiles. Meanwhile, a reduction of the growth rate occurs due to: (1) metabolic burden caused by algU regulator overexpression; (2) altered nitrogen metabolism caused by mutations in rpoN and caused by changes in the RpoN regulon transcriptional profiles; (3) altered DNA supercoiling caused by gyrA gene mutations; (4) altered DNA replication caused by mutations on important component of the DNA polymerase ( lig , dnaG , holC ); (5) altered transcription caused by rpoB gene mutations; and (6) altered translation caused by mutations in ribosomal proteins genes. Such reduction in growth rate determines an increased tolerance to antibiotics which finally helps P. aeruginosa to persist in the CF lungs.

Article Snippet: A total of 1 μg of RNA from samples with a RIN higher than 9 were used for preparing strand-specific sequencing libraries using the KAPA RNA HyperPrep Kit (Roche) after rRNA depletion obtained using P. aeruginosa riboPOOL kit (siTOOLs Biotech).

Techniques: Gene Expression, Over Expression