Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Schematic graph presenting the crucial role of proprotein convertase subtilisin/kexin type 9 (PCSK9) on LDL receptor turnover in hepatocytes (created with BioRender.com , accessed on 15 November 2023).

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Integrated optical density (IOD) of DNA bands after conventional reverse transcription PCR (RT–PCR) ( B ) and agarose gel electrophoresis ( A ). An equal amount of RNA taken from the T7 RNA polymerase reaction served as a template. The control sample contained a PNA oligomer that was not complementary to the PCSK9 coding sequence. This sample was arbitrarily assigned a value of 100%. In Panel ( B ), dashed line bars represent real-time PCR, while white dotted black bars represent conventional PCR after 15 cycles, in comparison to the control PNA. *— p < 0.05 vs. ctrlPNA for RT-PCR, #— p < 0.05 vs. ctrlPNA for conventional PCR.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Sequencing, Real-time Polymerase Chain Reaction, Comparison

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Western blot image of an in vitro transcription/translation reaction product resolved by electrophoresis with appropriate integrated optical density (I.O.D) values ( A ) and the relative amount of protein with respect to control sample ( B ). 1–4: Samples that contained InitPNA, Ex1PNA, Ex2PNA, and control (non-PCSK9 specific) PNA, respectively. M: Color Prestained Protein Standard, Broad Range (New England Biolabs, no. Cat. P7712). The PCSK9-specific band is depicted with an arrow. A total amount of 10 µg (25 µL sample volume) of purified products from the transcription/translation reaction was loaded onto the gel. The initial protein concentration in the samples was assessed spectrophotometrically. Measurements were made using ImageJ 1.53t software [ , ] *: p < 0.05 vs. control.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Western Blot, In Vitro, Electrophoresis, Control, Purification, Protein Concentration, Software

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: The representative HPLC traces of GFP-tagged PCSK9 synthetic protein after addition of PNA oligomers ( A ). Area under the curve (AUC) values were calculated for each sample (run in triplicates) from chromatography peaks of GFP-tagged PCSK9 protein synthesized with the presence of different PNA oligomers ( B ) *: p < 0.05 vs. CtrlPNA.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Chromatography, Synthesized

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Sequences of synthesized PNA-Ahx-NLS conjugates (Ahx-6-aminohexanoic acid).

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Synthesized, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Properties of PNA oligomers used in this study.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Sequencing, Control

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: High-performance liquid chromatography (HPLC) conditions for analysis of GFP-tagged PCSK9 protein after cell-free synthesis in vitro. The reaction was carried out under the following conditions: flow: 0.8 mL/min; detection (ex/em): 482/511 nm; column temperature: 40 °C; injection volume: 10 µL; mobile phase A: 0.1% trifluoroacetic acid in deionized water; mobile phase B: 0.1% trifluoroacetic acid in acetonitrile.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: High Performance Liquid Chromatography, In Vitro, Injection