antisense oligonucleotides  (Ribobio co)

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: LINE1, cardiac senescence and genomic instability respond to MI in mice. (A-B) RT‒qPCR analysis showing the expression of ORF1 and ORF2 of LINE1 in the zone, peri-infarcted zone and infarcted core of ischemic ventricular tissue on the indicated days after MI (n = 4). (C) A representative Western blot analysis revealed that the level of the DNA damage marker p-H2A.X Ser139 and the epigenetic markers H3K4me3, H3K9me3, and H3K27me3 in the heart on the indicated days after MI. (D) A representative Western blot analysis confirmed the expression of the LINE1 retrotransposon and p53 and p21 in the heart at the indicated days after MI. ( E ) Quantitative data of the Western blot results (n = 3). All the data are expressed as the mean ± s.d. Significant differences were determined via one-way ANOVA with Tukey's HSD multiple comparison post hoc test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: Expressing, Western Blot, Marker, Comparison

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: Preparation and characterization of engineered endothelium-targeting and LINE1 ASO-packaged EVs. (A) Schematic illustration of the preparation of DSPE-PEG-SBP. (B) Schematic illustration of the fabrication of SBP-LINE1-EVs through internal and external modification by SBP anchoring and LINE1-ASO electroporation. (C) Nanoparticle tracking analysis (NTA) was used to analyze the particle size distributions of EVs, SBP-EVs and SBP-LINE1-EVs. (D) Transmission electron microscopy (TEM) images of EVs and SBP-EVs. Scale bars, 100 nm. (E) Western blot analysis of the EV markers ALIX, TSG101, CD9 and CD63 in EVs, SBP-EVs, SBP-LINE1-EVs and hP-MSCs. All the data are expressed as the mean ± s.d. P values were calculated via one-way ANOVA with Tukey's HSD multiple comparison post hoc test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: Modification, Electroporation, Transmission Assay, Electron Microscopy, Western Blot, Comparison

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: In vitro assessment of the cargo transfer capacity and therapeutic potential of SBP-LINE1-EVs. (A) Experimental design for assessing the permeation effect of SBP-EVs through HUVEC monolayers. (B-D) Representative images of DiI-labeled EVs, DiI-labeled SBP-EVs and α-Actinin immunofluorescence staining of CMs in the lower chamber under normoxic conditions (B) and OGD-induced stress conditions (C) . The intensity of DiI signals in the CMs was quantified (n = 6) (D) . An anti-CD62E antibody and/or anti-CD62P antibody was added to the upper chamber of the hypoxic system for 2 h to block CD62E and/or CD62P in HUVECs after pretreatment with OGD. Scale bar, 50 μm. (E) Immunofluorescence staining of 5-FAM-labeled LINE1-ASO delivery and α-Actinin. FAM-labeled LINE1-ASO was electroporated into EVs, and FAM-LINE1-ASO-containing SBP-EVs were added to the upper chamber of the transwell and incubated for 6 h. Scale bar, 50 μm. (F) The degree of CM senescence was analyzed via β-gal staining after the internalization of SBP-LINE1-EVs, which penetrated from the upper chamber to the lower chamber of the transwell. β-gal-positive CMs under different conditions were quantified (n = 3). Scale bar, 100 μm. (G) The expression level of LINE1 in CMs was analyzed via RT‒qPCR (n = 4). (H) Representative images of β-gal-stained CMs after treatment with Scrambled ASO-packaged SBP-EVs or LINE1-ASO-packaged SBP-EVs for 24 h. Scale bar, 100 μm; n = 3. All the data are expressed as the mean ± s.d. P values were calculated via one-way ANOVA with Tukey's HSD multiple comparison post hoc test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: In Vitro, Labeling, Immunofluorescence, Staining, Blocking Assay, Incubation, Expressing, Comparison

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: SBP-LINE1-EVs improved myocardial infarction injury and reduced adverse cardiac remodeling in mice. (A) Experimental schedule showing the study design of the treatments for EVs, SBP-EVs and SBP-LINE1-EVs. (B) TTC staining images and quantification were performed to evaluate the infarct area of ischemic ventricle tissue via vein injection of PBS, EVs, SBP-EVs or SBP-LINE1-EVs (n = 4). Scale bar, 2 mm. (C-D) Serum levels of creatine kinase-myocardial band isoenzyme (CK-MB) ( C ) and lactate dehydrogenase (LDH) ( D ) in mice treated with EVs, SBP-EVs or SBP-LINE1-EVs on day 4 (n ≥ 12). (E) Representative images of M-mode echocardiography. Statistical analysis of cardiac function revealed significant improvements in the left ventricular ejection fraction (EF), fractional shortening (FS), left ventricular internal diameter at end diastole (LVIDd), and left ventricular internal diameter in the sole (LVIDs) on days 7 and 14 (n = 5). (F) Representative images of Masson's trichrome staining on day 28 after MI. The infarct size of the left ventricle was quantified via Masson's trichrome staining (n = 5). Scale bar, 1 mm. (G) Western blot images and quantitative data of the expression of cardiac fibrosis-related genes, including α-SMA, COL1A1, and COL3A1, in ventricle tissue from each group of mice on day 28 after MI (n = 3). All the data are expressed as the mean ± s.d. P values were calculated via one-way ANOVA with Tukey's HSD multiple comparison post hoc test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: Staining, Injection, Western Blot, Expressing, Comparison

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: SBP-LINE1-EVs relieved cardiac senescence and the innate immune response by maintaining genomic stability and inhibiting the cGAS-STING signaling pathway. (A) Representative images of LINE1 and the cardiac marker α-SA immunofluorescence staining of ventricle tissue in each group of mice at 28 days after MI . The expression of LINE1 was quantified (n = 5). Scale bar, 50 μm. (B) Representative images of β-gal-stained samples from each group of mice at 28 days after MI (n = 5). (C-D) Representative images of immunofluorescence staining for the cytoplasmic DNA sensors cGAS (C) and STING (D) in ventricular tissue from each group of mice 28 days after MI. The expression of cGAS and STING was quantified (n = 5). Scale bar, 50 μm. (E) Representative images of IFN-β immunofluorescence staining of ventricle tissue in each group of mice 28 days after MI , and the expression of IFN-β was quantified (n = 5). Scale bar, 100 μm. (F) Western blot images of LINE1, cGAS, and STING and their downstream phosphorylated forms of TBK1, phosphorylated forms of IRF3, and IFN-β, and the DNA damage marker p53 together with the senescence marker p21 in ventricle tissue from each group of mice at 28 days after MI (n = 3). (G) Western blot images of the DNA damage marker p-H2A.X Ser139 and the epigenetic markers H3K4me3, H3K9me3, and H3K27me3 in the ventricle tissue of each group of mice at 28 days after MI (n = 3). All the data are expressed as the mean ± s.d. P values were calculated via one-way ANOVA with Tukey's HSD multiple comparison post hoc test. ∗ P < 0.05, ∗∗ P < 0.01, n ≥ 6.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: Marker, Immunofluorescence, Staining, Expressing, Western Blot, Comparison

Journal: Bioactive Materials

Article Title: Delivering LINE1 antisense oligonucleotides via endothelial targeting extracellular vesicles to ameliorate myocardial infarction-induced cardiac senescence

doi: 10.1016/j.bioactmat.2025.07.008

Figure Lengend Snippet: Schematic diagram depicting the therapeutic effects of intravenous administration of SBP-LINE-EVs for MI treatment. SBP-LINE1-EVs exhibit a selective targeting tendency to the injured heart with an enhanced myocardial protection effect by LINE1-ASO delivery. SBP-LINE1-EVs protected against myocardial senescence by inhibiting reactivation of the innate immune cGAS-STING-TBK1-IRF3 pathway and further repressing the expression of senescence-associated secretory phenotype (SASP) factors.

Article Snippet: To generate the EVs containing LINE1-ASO, 500 μg of naive EVs and 100 or 200 pmol of LINE1 antisense oligonucleotide (LINE1-ASO) were diluted in Gene Pulser Electroporation Buffer and transferred to ice-cold 0.4 cm or 0.2 cm Gene Pulser/MicroPulser Electroporation Cuvette (Bio-Rad, Hercules, CA, USA).

Techniques: Expressing