Synthesis of an <i>Ortho</i>‐Functionalized Tetrafluorinated Azobenzene Phosphoramidite for Incorporation into a Tetrafluorinated Azobenzene‐Containing siRNA for Photocontrolled Gene Silencing
Bibliographic record
Abstract
Abstract This article presents the detailed synthesis and characterization protocols for an ortho‐functionalized tetrafluorinated azobenzene containing siRNA, which has photoswitchable properties. To design this tetrafluorinated azobenzene scaffold, several synthetic steps are performed to generate a symmetrical tetrafluorinated azobenzene diol. This diol is treated with dimethoxytrityl chloride (DMT‐Cl) to protect one of the alcohols. Next, the DMT‐protected tetrafluorinated monoalcohol is phosphitylated to afford the DMT‐phosphoramidite building block used for solid‐phase synthesis. This paper also contains the detailed biophysical characterization, biological testing, and photo‐switching protocols of an ortho‐functionalized fluorinated azobenzene containing siRNA (F‐siRNA), which has photoswitchable properties that can be controlled with visible light. First, the F‐siRNA was characterized by annealing the sense and antisense strands together and then measuring the circular dichroism (CD) profile and melting temperature (Tm) of the duplexes. Second, biological testing of the F‐siRNA is performed in cell culture to determine their gene silencing efficacy. Finally, their gene‐silencing activities are measured after exposure to green light to inactivate the F‐siRNA, followed by blue light, which reactivates the F‐siRNA. The F‐siRNA can be kept inactive for up to 72 hr and reactivated at any time within this 72‐hr window. © 2023 Wiley Periodicals LLC.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| 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) | 0.001 | 0.001 |
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 source (direct Gemma or distilled Codex), 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".