592. A Comparison of Length on the Potency and Toxicity of Small Interfering and Short Hairpin RNAs Targeting a Highly Conserved Site in HIV-1 RNA Coding for the Gag Polyprotein
Notice bibliographique
Résumé
Small interfering RNAs (siRNAs) and their precursors, short hairpin RNAs (shRNAs), are being developed for the treatment of HIV-1 infection as drug and gene therapies, respectively. We have previously identified a target site in HIV-1 RNA coding for the Gag polyprotein that was highly conserved across diverse HIV-1 genomes and particularly accessible to a ribozyme. An shRNA targeting this site was also shown to be a potent inhibitor of HIV-1 production from diverse strains. Several studies have shown that the gene silencing effects of siRNAs and shRNAs are highly dependent on their lengths. While optimal lengths have been proposed, conflicting results suggest that empirical data is required to identify the best format for any particular target site. Using the target site identified in our previous study, we designed siRNAs and shRNAs with different lengths to identify the optimal therapeutic format for further development. Symmetrical and asymmetrical siRNAs, with intended sense-strand lengths of 17 to 29 bases, showed a similar trend in potency against HIV-1 production and the most effective molecules had sense-strand lengths of 27 and 29 bases. In contrast, shRNAs with stem lengths of 20 and 21 base pairs were most effective, with a gradual decrease in potency as the stem length was increased to 29 base pairs. To compare potential immune stimulation and toxicity of the different siRNAs designed, cellular proliferation and phosphorylation of protein kinase R (PKR) were evaluated in HEK293T and MCF7 cells. While phosphorylation of PKR was not detected, minor effects on cell proliferation were observed for some siRNAs at concentrations several fold higher compared to their minimum inhibitory concentrations against HIV-1 production in HEK293T cells. Our results are consistent with reports suggesting that longer siRNAs are more potent compared to their sequence matched canonical formats; however, a standard longer siRNA format, with a 25 base pair sense-strand, was not long enough to observe an increase in potency over canonical formats. Our results also suggest that for certain target sites, there is no correlation between the optimal length of siRNAs and shRNAs. Overall, our results highlight the importance of obtaining empirical data on the optimal si/shRNA format for any particular target site, and provide a set of optimized molecules targeting the Gag coding sequence of HIV-1 RNA for further development.
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».