Successful inhibition of Dnmt1 expression in bovine using RNA interference.
Bibliographic record
Abstract
Epigenetic modifications, such as methylation are essential for normal mammalian development. Dnmt1 is the predominant and most active methylase, which is responsible for copying methylation patterns after replication. During early development, Dnmt1 is highly regulated to support passive demethylation and normal reprogramming. In somatic cells, Dnmt1 is localized in the nucleus and its introduction into cloned embryos is believed to be the main limiting factor for proper reprogramming of the donor genome. The aim of this study was to transiently suppress Dnmt1 by RNA interference (RNAi) to investigate the role of Dnmt1 during early development and its effect on nuclear transfer technology. RNA and protein extracts were collected from fibroblasts treated with control or Dnmt1 RNAi. Real time RT‐PCR revealed a 93.4% reduction of Dnmt1 transcript levels. Consistently, Western blot analysis showed a protein decrease of up to 98.8%. Gene expression of housekeeping genes, GAPD, ACTB and lamin A/C, were not affected after RNAi treatment however, expression of HPRT1, H19, MMP1 and SCYA genes was increased. This is the first report to demonstrate a specific and highly effective knockdown of bovine Dnmt1 by RNAi at both mRNA and protein levels. Dnmt1 RNAi could be a useful tool to better understand epigenetic reprogramming events as well as syndromes related to abnormal methylation patterns. (Funding by CONACyT & NSERC)
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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".