423 PRODUCTION OF CLONED PIGS EXPRESSING APOLIPOPROTEIN E-SPECIFIC SMALL HAIRPIN (shRNA)
Notice bibliographique
Résumé
Apolipoprotein E (apo E) is a known risk factor for developing premature atherosclerosis and Alzheimer’s syndrome. The aim of this study was to create a pig model with reduced apo E levels using RNA interference (RNAi) and somatic cell nuclear transfer (SCNT) technologies. Three synthetic small interfering RNA targeting the porcine apo E mRNA were designed, and the knockdown efficiency was assessed in cultured porcine granulosa cells by real-time PCR. The observed apo E knockdown efficiency ranged from 45 to 82% compared with control cells, indicating the targeted degradation of apoE mRNA.A small hairpin RNA (shRNA) expressing vector was constructed in PRNAT.U6.Neo (Genscript Corp., Piscataway, NJ, USA) based on the most effective apo E RNAi sequence under the control of polymerase III (U6) promoter, and then introduced into fetal porcine fibroblast cells. Clones were selected by neomycin treatment and green fluorescent protein (GFP) expression. SCNT was performed using IVM oocytes collected from prepubertal gilts. Oocyte maturation and activation and embryo culture were performed as previously described (Nascimento et al. 2009 Reprod. Domest. Anim. in press). Embryos were cultured in vitro for 5 to 6 days, briefly exposed to fluorescent light to confirm GFP expression, and then surgically transferred into the uterus of recipient gilts. The recipient gilts were synchronized by daily oral administration of altrenogest (20 mg day-1; Regu-Mate®, Intervet, Millsboro, MD, USA) for 12 or 13 days, followed by 1000 IU of eCG injected in the last day of altrenogest treatment and 500 IU of hCG 72 h later. Pregnancy diagnosis was performed by ultrasonography at Day 20 to 25 after embryo transfer, and parturition was induced by injecting PGF2? (10 mg of dinoprost tromethamine; Lutalyse®, Pfizer Canada Inc., Montreal, QC, Canada) at Day 115 of pregnancy. Rates of cleavage (74.7%) and development to the blastocyst stage (37.2%) were comparable with that of embryos reconstructed with nontransfected cells from the same cell line. A total of 309 embryos were transferred to 5 recipients, of which 3 became pregnant and farrowed. Seven live and 1 stillborn piglets were delivered naturally. The presence of the introduced plasmid and the expression of the GFP transgene tag were confirmed by PCR in placental and umbilical tissues of all the piglets. Six cloned pigs have survived after weaning and exhibit no obvious morphological defects. The status of apo E gene expression is currently under investigation. Supported by a NSERC Discovery Grant to VB.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
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 | 0,001 | 0,001 |
| 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 ».