研究目的
To improve the cellular uptake and stability of exogenous nucleic acids by employing nanosized particles consisting of oligonucleotides, specifically BODIPY-labeled oligonucleotides, for efficient gene regulation in living cells and tumor tissues.
研究成果
The study demonstrated that BODIPY-labeled oligonucleotides form aggregates that efficiently penetrate cell membranes via scavenger-receptor-mediated endocytosis, leading to effective gene regulation in living cells and tumor tissues. This simple modification offers a promising approach for genetic manipulation without the need for complex additives or invasive techniques.
研究不足
The study focused on a specific modification (BODIPY-labeled oligonucleotides) and its effects on cellular uptake and gene regulation. The comparison with conventional transfection systems indicated that the present system is less efficient in suppressing specific gene activity, highlighting a limitation in efficiency compared to existing methods.
1：Experimental Design and Method Selection:
The study utilized BODIPY-labeled oligonucleotides to form nanosized aggregates in aqueous solution. The aggregates' formation, cellular uptake, and gene regulation efficiency were evaluated.
2：Sample Selection and Data Sources:
Human cell line of lung adenocarcinoma (A549) and fibrosarcoma cell line (HT1080) were used for cellular experiments. Colon carcinoma cell line (colon 26-luc) was transplanted into nude mice for in vivo studies.
3：List of Experimental Equipment and Materials:
Confocal microscopy, spectrofluorophotometer, nanoparticle tracking analysis, and zetasizer were used. BODIPY-labeled oligonucleotides were purchased from Japan Bioservice and Eurofins genetics.
4：Experimental Procedures and Operational Workflow:
The formation of aggregates was monitored using fluorescence spectra. Cellular uptake was assessed via confocal microscopy, and gene regulation was evaluated by measuring luciferase activity.
5：Data Analysis Methods:
Fluorescence intensity was measured to quantify cellular uptake. Gene silencing efficiency was assessed by normalizing luciferase activities against protein content.
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