reporter mrna encoding enhanced green fluorescent protein (egfp)  (TriLink)

Journal: Frontiers in Pharmacology

Article Title: PepFect14 mediates the delivery of mRNA into human primary keratinocytes and in vivo

doi: 10.3389/fphar.2023.1219761

Figure Lengend Snippet: PF14-mRNA nanoparticles are with suitable characteristics for cellular delivery. ( A,B ) For DLS, nanoparticles were prepared using PF14 and four different mRNAs: mCherry, luciferase, EGFP and Cy5-mRNA at charge ratio (CR) 2:1. If indicated, nanoparticles were supplemented with PS80, chloroquine, MgCl 2 and/or CaCl 2 . The white bars represent the treatments, where only PF14 or indicated mRNAs were added to the cells. Data are representative of three technical replicates and expressed as ± SEM, n = 3, one-way ANOVA with post-hoc Šidák test was used, ns – not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: mRNAs: Anti-reverse cap analogue (ARCA) mRNA encoding enhanced green fluorescent protein (EGFP), i.e., EGFP-mRNA (5-moUTP), ARCA Cy5-labeled EGFP-mRNA (5mo-UTP), and CleanCap mCherry-mRNA (5mo-UTP) were purchased from APExBIO Technology (Houston, TX, USA), and CleanCap Fluc-mRNA, i.e., luciferase-mRNA (5moU) from TriLink Biotechnologies (San Diego, CA, USA).

Techniques: Luciferase

Journal: bioRxiv

Article Title: High throughput intracellular delivery by viscoelastic mechanoporation

doi: 10.1101/2023.04.24.538131

Figure Lengend Snippet: (a) eGFP expression 24 hours after mRNA transfection to Jurkat cells for a range of extracellular mRNA concentrations. (b) T cell receptor (TCR) expression in Jurkat cells 2 days after transfection of Cas9 RNP complexes designed to knock out the TCR, for a range of RNP concentrations ( n = 3 for experimental conditions, n = 1 for controls). (c) Viability and delivery of 70 kDa FITC-dextran to HEK293T cells, 24 hours after processing, for three delivery buffer compositions and 2 different chip pressures (Unstretched, n = 2 ; PBS, n = 1 ; Cytomix buffers, n = 3 ). (d) Viability and delivery of 70 kDa FITC-dextran to primary activated T cells, about 90 minutes after processing, for increasing chip pressures ( n = 2 ).

Article Snippet: For mRNA transfection, we used an mRNA encoding enhanced green fluorescent protein (eGFP) with ARCA cap modifications (Apexbio Technologies, Fisher Scientific #50-199-8310).

Techniques: Expressing, Transfection, Knock-Out

Journal: Pharmaceutics

Article Title: mRNA-Loaded Lipid Nanoparticles Targeting Dendritic Cells for Cancer Immunotherapy

doi: 10.3390/pharmaceutics14081572

Figure Lengend Snippet: Comparison of the A-11-LNPs with clinically relevant formulations. ( A ) Nluc expression in spleen and inguinal LN 24 h after intravenous administration of Nluc-mRNA-loaded formulations at a dose of 0.5 mg mRNA/kg. ( B ) Percentage of EGFP + splenic DCs 24 h after the intravenous administration of EGFP-mRNA-loaded formulations at a dose of 0.5 mg mRNA/kg. ( C ) I-A/I-E expression in splenic DCs 24 h after intravenous administration of EGFP-mRNA-loaded formulations at a dose of 0.5 mg mRNA/kg. n = 3. ** p < 0.01.

Article Snippet: The ovalbumin (OVA)-encoding mRNA (cat# L-7610) and enhanced green fluorescent protein (EGFP)-encoding mRNA (cat# L-7601) were purchased from Trilink BioTechnologies (San Diego, CA, USA).

Techniques: Expressing

Journal: Pharmaceutics

Article Title: mRNA-Loaded Lipid Nanoparticles Targeting Dendritic Cells for Cancer Immunotherapy

doi: 10.3390/pharmaceutics14081572

Figure Lengend Snippet: Prophylactic and therapeutic antitumor activity. ( A ) Prophylactic antitumor activity of the A-11-LNPs in E.G7-OVA tumor-bearing mice. The OVA mRNA-loaded A-11-LNPs were intravenously injected at the indicated doses twice on 14 and 7 days before tumor inoculation. n = 3–4. ( B ) The therapeutic antitumor activity of the A-11-LNPs, MC3-LNP, RNA-LPX, and B-8-LNPs. E.G7-OVA tumor-bearing mice were intravenously injected with OVA mRNA-loaded formulations at two doses of 0.03 mg mRNA/kg on day 8 and 11. n = 5. * p < 0.05, ** p < 0.01. ( C – E ) Expression of activation markers CD40 ( C ), CD80 ( D ), and CD86 ( E ) in splenic DCs 24 h after an intravenous injection of OVA mRNA-loaded formulations at a dose of 0.03 mg mRNA/kg. n = 3. * p < 0.05, ** p < 0.01. ( F – H ) Expression of an activation marker CD69 in splenic B cells ( F ), T cells ( G ), and NK cells ( H ) 24 h after an intravenous injection of OVA mRNA-loaded formulations at a dose of 0.03 mg mRNA/kg. n = 3. ** p < 0.01.

Article Snippet: The ovalbumin (OVA)-encoding mRNA (cat# L-7610) and enhanced green fluorescent protein (EGFP)-encoding mRNA (cat# L-7601) were purchased from Trilink BioTechnologies (San Diego, CA, USA).

Techniques: Activity Assay, Injection, Expressing, Activation Assay, Marker

Journal: Pharmaceutics

Article Title: mRNA-Loaded Lipid Nanoparticles Targeting Dendritic Cells for Cancer Immunotherapy

doi: 10.3390/pharmaceutics14081572

Figure Lengend Snippet: Safety of the A-11-LNPs. Serum chemistry parameters, ALT ( A ), AST ( B ), T-BIL ( C ), LDH ( D ), BUN ( E ), and CRE ( F ) were measured 24 h after the last injection of OVA mRNA-loaded A-11-LNPs at two doses of 0.03 mg mRNA/kg. n = 3.

Article Snippet: The ovalbumin (OVA)-encoding mRNA (cat# L-7610) and enhanced green fluorescent protein (EGFP)-encoding mRNA (cat# L-7601) were purchased from Trilink BioTechnologies (San Diego, CA, USA).

Techniques: Injection