dynabeads mrna direct micro kit  (Thermo Fisher)

Journal: iScience

Article Title: A key role for the exoribonuclease XRN1 in regulating the hepatitis B viral transcriptome

doi: 10.1016/j.isci.2026.116328

Figure Lengend Snippet: XRN1 regulation of the HBV transcriptome (A) HBV transcript mapping. Cartoon depicting the genomic viral RNA with the 5′ and 3′ epsilon stem loops, canonical transcription start sites, and major viral RNAs along with open reading frames (core, Pol, L, M, S, and X). HepG2-NTCP WT or XRN1 KO22 cells were infected with HBV, RNAs extracted at 6 dpi, and analyzed by long-read sequencing. Read counts for pC, pg, preS1, preS2, S, and X transcripts (left), along with spliced isoforms (right), are shown. Data are presented for 6 independent samples and statistical significance assessed using unpaired t tests (corrected for multiple comparisons, ∗ p < 0.05). (B) In vitro pgRNA digestion by XRN1. Capped-analog modified mRNA was synthesized by in vitro transcription. The capped and decapped pgRNA was incubated with XRN1, evaluated by agarose gel electrophoresis, and northern blotting using an HBV probe. Please see also .

Article Snippet: In vitro pgRNA transcription was performed in a 20 μL final volume using a T7 mScript standard mRNA production system by following the protocol provided by the manufacturer (CellScript), the DNA template was digested with 10 μL of a DNase cocktail containing 10 U of DNase I (NEB), 3 μL 10 X DNase I buffer, 2 μL nuclease-free water and incubated at 37°C for 20 min. After purification of the RNA using RNeasy kit (Qiagen), capping and polyadenylation were performed following Cap1 and Cap0 mRNA protocol described in the user’s manual, and the RNA was again purified using RNeasy kit.

Techniques: Infection, Sequencing, In Vitro, Modification, Synthesized, Incubation, Agarose Gel Electrophoresis, Northern Blot

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Schematic illustration of in vivo tumor immunotherapy enhanced by mRNA/HNPs through intravenous injection. H18 lipid, DOPE, cholesterol, DMG-PEG 2000 and mRNA were mixed to form mRNA/H 18 NPs with the special multilamellar concentric nanostructures. Following intravenous administration, mRNA/H 18 NPs demonstrated preferential adsorption of complement C3 proteins to form a characteristic protein corona, resulting in specific mRNA transfection in the spleen, especially in splenic dendritic cells. When encapsulating tumor antigen-encoding mRNA, the mRNA/H 18 NPs achieved precise transfection of the antigen mRNA in splenic dendritic cells. This targeted delivery stimulated dendritic cell maturation and subsequent antigen presentation, initiating robust T cell priming. The activated antigen-specific cytotoxic T lymphocytes then infiltrated into tumor tissues, ultimately inducing tumor cell elimination.

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Injection, Adsorption, Transfection, Immunopeptidomics

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Preparation and Characterization of Optimal mRNA/H 18 NPs. (A) Schematic illustration of mRNA/H 18 NPs preparation. (B) The size distribution and (C) zeta potential of optimized mRNA/H 18 NPs. (D) The apparent p K a of mRNA/H 18 NPs. (E) Cryo-EM image of optimized mRNA/H 18 NPs. Scale bar = 50 nm. (F) Representative image and percentage of bioluminescence in major organs of mice following intravenous injection of mLuc/H 18 NPs. (G)-(I) Stability test for mRNA/H 18 NPs. (G) Size and PDI of mRNA/H 18 NPs when stored at 4 °C for different days (0, 3, 5, 7). (H) Left: Total bioluminescence flux in the spleen of mice 6 h after intravenous injection of mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). Right: Representative bioluminescence images of major organs of mice 6 h after intravenous injection of different mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). (I) Size and PDI of mRNA/H 18 NPs when diluted with PBS by different times. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: Zeta Potential Analyzer, Cryo-EM Sample Prep, Injection

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo splenic DC-specific transfection of mRNA/H 18 NPs and in vitro protein corona analysis of mRNA/H 18 NPs. (A) EGFP protein expression in splenic cell subsets of C57BL/6J mice 24 h post intravenous injection of different formulations. (B) The top 5 most abundant plasma proteins adsorbed on mRNA/H 18 NPs (C3: Complement C3; Ighm: Immunoglobulin heavy constant mu; Hbat1: Alpha-globin; Itih4: Inter alpha-trypsin inhibitor, heavy chain 4; Cnn2: Calponin). (C) Heatmap plot of major proteins in the protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. PBS group was used as a negative control. (D) Quantification of major adsorbed protein categories of different formulations. (E) Complement C3 abundance in protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. (F) Bioluminescence images of major organs and (G) Quantification of total bioluminescence flux in the spleen from C57BL/6J mice 6 h after intravenous injection of mLuc/H 18 NPs (mLuc dose of 0.25 mg kg −1 ). Mice were pre-treated with cobra venom factor (CVF) or PBS. (H) Fluorescence quantification of Cy5 mRNA delivered by uncoated or complement C3-coated Cy5-mRNA/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. (I) Bioluminescence intensity of luciferase protein translated from mRNA delivered by uncoated or complement C3-coated mLuc/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Transfection, In Vitro, Expressing, Injection, Clinical Proteomics, Negative Control, Combined Bisulfite Restriction Analysis Assay, Fluorescence, Incubation, Blocking Assay, Luciferase

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Schematic illustration of in vivo tumor immunotherapy enhanced by mRNA/HNPs through intravenous injection. H18 lipid, DOPE, cholesterol, DMG-PEG 2000 and mRNA were mixed to form mRNA/H 18 NPs with the special multilamellar concentric nanostructures. Following intravenous administration, mRNA/H 18 NPs demonstrated preferential adsorption of complement C3 proteins to form a characteristic protein corona, resulting in specific mRNA transfection in the spleen, especially in splenic dendritic cells. When encapsulating tumor antigen-encoding mRNA, the mRNA/H 18 NPs achieved precise transfection of the antigen mRNA in splenic dendritic cells. This targeted delivery stimulated dendritic cell maturation and subsequent antigen presentation, initiating robust T cell priming. The activated antigen-specific cytotoxic T lymphocytes then infiltrated into tumor tissues, ultimately inducing tumor cell elimination.

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Injection, Adsorption, Transfection, Immunopeptidomics

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Preparation and Characterization of Optimal mRNA/H 18 NPs. (A) Schematic illustration of mRNA/H 18 NPs preparation. (B) The size distribution and (C) zeta potential of optimized mRNA/H 18 NPs. (D) The apparent p K a of mRNA/H 18 NPs. (E) Cryo-EM image of optimized mRNA/H 18 NPs. Scale bar = 50 nm. (F) Representative image and percentage of bioluminescence in major organs of mice following intravenous injection of mLuc/H 18 NPs. (G)-(I) Stability test for mRNA/H 18 NPs. (G) Size and PDI of mRNA/H 18 NPs when stored at 4 °C for different days (0, 3, 5, 7). (H) Left: Total bioluminescence flux in the spleen of mice 6 h after intravenous injection of mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). Right: Representative bioluminescence images of major organs of mice 6 h after intravenous injection of different mLuc/H 18 NPs stored at 4 °C for different days (0, 3, 5, 7). (I) Size and PDI of mRNA/H 18 NPs when diluted with PBS by different times. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: Zeta Potential Analyzer, Cryo-EM Sample Prep, Injection

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo splenic DC-specific transfection of mRNA/H 18 NPs and in vitro protein corona analysis of mRNA/H 18 NPs. (A) EGFP protein expression in splenic cell subsets of C57BL/6J mice 24 h post intravenous injection of different formulations. (B) The top 5 most abundant plasma proteins adsorbed on mRNA/H 18 NPs (C3: Complement C3; Ighm: Immunoglobulin heavy constant mu; Hbat1: Alpha-globin; Itih4: Inter alpha-trypsin inhibitor, heavy chain 4; Cnn2: Calponin). (C) Heatmap plot of major proteins in the protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. PBS group was used as a negative control. (D) Quantification of major adsorbed protein categories of different formulations. (E) Complement C3 abundance in protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. (F) Bioluminescence images of major organs and (G) Quantification of total bioluminescence flux in the spleen from C57BL/6J mice 6 h after intravenous injection of mLuc/H 18 NPs (mLuc dose of 0.25 mg kg −1 ). Mice were pre-treated with cobra venom factor (CVF) or PBS. (H) Fluorescence quantification of Cy5 mRNA delivered by uncoated or complement C3-coated Cy5-mRNA/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. (I) Bioluminescence intensity of luciferase protein translated from mRNA delivered by uncoated or complement C3-coated mLuc/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. Data were shown as mean ± SD (n = 3).

Article Snippet: Luciferase mRNA, OVA mRNA, and EGFP mRNA were purchased from Novoprotein (Suzhou, China).

Techniques: In Vivo, Transfection, In Vitro, Expressing, Injection, Clinical Proteomics, Negative Control, Combined Bisulfite Restriction Analysis Assay, Fluorescence, Incubation, Blocking Assay, Luciferase