44 POSSIBILITY OF PRODUCTION OF A TRANSGENIC CLONED COW USING THE SOMATIC CELL NUCLEAR TRANSFER OF HUMAN ALPHA 1-ANTITRYPSIN GENE-TRANSFECTED CELLS
Notice bibliographique
Résumé
The present study was designed to produce, using a somatic cell nuclear transfer (SCNT), transgenic cloned cows that secreted a therapeutic protein into milk. Bovine cumulus cells were transfected with a plasmid that contained the alpha 1-antytrypsin DNA and green fluorescent protein (GFP) reporter gene using a Fugene 6 as a lipid carrier. In vitro-matured bovine oocytes were enucleated followed by confirmation of enucleation via nuclear staining. The GFP-expressing donor cells with the transgene were selected under blue fluorescent light just before injection of cells. The SCNT was performed according to the established system in our laboratory (Jang G et al., 2003 Reprod. Fertil. Dev. 15, 179–185). The couplets were fused, chemically activated, and cultured in modified synthetic oviduct fluid (mSOF) at 39°C in a humidified atmosphere of 5% CO2 in air for up to 7 days. The developmental ability (in Experiment 1) and number of inner cell mass (ICM) and trophectoderm (TE) cells (in Experiment 2) in transgenic and non-transgenic cloned embryos determined by differential staining as described by Thouas et al., (2001 Reprod. Biomed Online 3, 35–29) were compared. In Experiment 3, only GFP-expressing blastocysts were transferred to the uteri of recipient cows. A general linear model consisting of ANOVA and a least squares method in the SAS program was used for statical analysis. The percentage of blastocysts was lower (P < 0.05) in transgenic cloned embryos compared to non-transgenic embryos (34% v. 20%). No difference (P < 0.05) in the cell number of ICM and TE cells between two groups of embryos was observed. Out of 47 recipient cows, three pregnancies were detected by non-return to estrus and by rectal palpation. However, the pregnancies were not maintained to term; 2 fetuses were aborted at Days 60, and 150, and one fetus was aborted at Day 240. The genomic DNA from the fetus aborted at Day 240 was prepared and analyzed for the expression of GFP and alpha 1-antitrypsin gene by polymerase chain reaction. The expected PCR products (710 bp for alpha 1-antytrypsin DNA and 431 bp for GFP) were obtained and confirmed by sequence analysis. In conclusion, the present study demonstrated that developmental competence of cloned embryos derived from transgenic donor cells was lower compared with cloned embryos derived from non-transfected donor cells. Although we failed to produce a viable transgenic cloned calf, expression of alpha 1-antitrypsin in fetal tissue represented the possibility of production of a transgenic cloned cow by SCNT. This study was supported by Biogreen 21-1000520030100000.
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 | 0,000 |
| 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 ».