1：Experimental Design and Method Selection:

The study designed a controllable RNA enrichment and releasing protocol using RNA aptamer-protein interactions and reversible light-inducible protein-protein interaction modules. Plasmids were constructed for fusion proteins and RNA fragments, and a recombinant 293T cell line was generated to produce engineered exosomes. Blue light was used to induce interactions for RNA enrichment on the plasma membrane, followed by exosome biogenesis and targeted delivery using AS1411 aptamers.

2：Sample Selection and Data Sources:

Human embryonic kidney 293T cells and human leukemia K562 cells were used. Samples included transfected cells, isolated exosomes, and treated cells for functional assays.

3：List of Experimental Equipment and Materials:

Equipment included confocal microscope (OLYMPUS FV1000), transmission electron microscope (Tecnai Spirit, FEI), flow cytometer (FACS Calibur, BD), qPCR instrument (Applied Biosystems 7300), and SIM microscope (Applied Precision V3). Materials included plasmids (e.g., from Addgene), Lipofectamine 3000, antibodies (e.g., anti-CD63, anti-PTEN), and kits for RNA extraction and cDNA synthesis.

4：3). Materials included plasmids (e.g., from Addgene), Lipofectamine 3000, antibodies (e.g., anti-CD63, anti-PTEN), and kits for RNA extraction and cDNA synthesis. Experimental Procedures and Operational Workflow:

4. Experimental Procedures and Operational Workflow: Cells were cultured and transfected with plasmids. Exosomes were isolated via ultracentrifugation, characterized using TEM, Western blot, and flow cytometry. RNA loading was quantified by qPCR. Targeted delivery was assessed by incubating exosomes with K562 cells, and functional effects were evaluated through apoptosis assays and Western blot for PTEN expression.

5：Data Analysis Methods:

Data were analyzed using ImageJ for Western blot quantification, flow cytometry for cell uptake and apoptosis, and qPCR with the 2?ΔΔCT method for RNA quantification. Statistical analysis included one-way ANOVA.