399. Solid Nanobeads Prevents Polycations from Entering the Nuclei and Enhanced Transfection Efficiency
Bibliographic record
Abstract
Objective: Employment of polycations for gene delivery to treat human diseases remains in doubt as long as the potential for polycations to adversely interact with host genes exists, which is incurred from entering of polycations into nuclei during gene delivery. In this study, we address this issue by conjugating polymer vector- gene complexes to silica nanobeads (SNB) and evaluated the transfection efficacy of marker and therapeutic genes. Methods: The poly-ethyleneimine vector (PEI)/DNA complexes were conjugated to SNB via a Sulfo-NHS-LC-Biotin linker. Luciferase and LacZ were used as reporter genes and VEGF-165 was used as therapeutic gene. Fluorescent labeling and confocal microscopy were employed to tack the paths of PEI/DNA complexes and SNB/PEI/ DNA complexes from endocytosis to gene expression. Stability of the SNB/PEI/DNA complexes was evaluated by DNase I protection assay. In vitro transfection efficacy of SNB/PEI/DNA was assessed in 4 different cell lines and compared with unconjugated PEI/DNA. In vivo, SNB/PEI/DNA complexes were systematically injected via tail vein of the mouse (n=12). Transfection efficacy of marker and therapeutic genes was assessed in the organs (heart, lung, spleen, kidney and liver) by luciferase assay, immunostaining and Western blotting. Results/Discussion: The SNB/PEI/DNA complexes were very stable and could provide more effective protection to DNA from nuclease degradation than PEI/DNA complexes. Confocal microscopy showed that PEI/DNA complexes entered nuclei in the form of ordered structures. In contrast, SNB/PEI/DNA complexes released DNA into the nuclei while SNB kept the PEI perinuclei of the cells. In vitro, 10- to 50-fold higher transfection efficiency for SNB/PEI/DNA complexes was observed compared with PEI/DNA complexes alone. In vivo, systematically injected SNB/PEI/DNA complexes resulted in effective over expression of marker and therapeutic genes. Conclusions: Conjugating polycation vector-gene complexes to solid nanobeads enhanced transfection efficiency and inhibited polymer to enter the nuclei, which may offer a novel way to prevent the risk of interference of polymer with native host DNA.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".