In vitro methods for the induction and maintenance of plant...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/00 (2006.01) C12N 5/04 (2006.01) C12N 15/82 (2006.01)

Patent

CA 2673663

The subject invention provides simple and consistent methods to break suspension cell aggregates to single cells with intact primary cell walls. The subject invention relates in part to cell separation of suspension cell aggregates cultured in medium containing pectin- degrading enzymes or tubulin de-polymerizing compounds including colchicine. The subject invention also relates to novel uses of compounds for such purposes. Another aspect of the subject invention relates to transformation of the subject, isolated cells. Such processes simplify and integrate single-cell-based transformation and selection processes into transgenic and transplastomic event-generation work processes. The subject invention also removes technical constraints and produces marker- free and uniformly expressing transgenic lines in a high throughput fashion to support various needs of animal health, biopharma, and trait and crop protection platforms.

L'invention concerne des procédés simples et cohérents pour la rupture d'agrégats de cellules en suspension en cellules uniques avec des parois cellulaires primaires intactes. L'invention concerne en partie la séparation de cellules d'agrégats de cellules en suspension cultivées dans un milieu contenant des enzymes dégradant la pectine ou des composés de dépolymérisation de la tubuline comprenant de la colchicine. L'invention concerne également de nouvelles utilisations de composés à de telles fins. Un autre aspect de l'invention concerne la transformation des cellules isolées du sujet. De tels procédés simplifient et intègrent des procédés de sélection et de transformation à base de cellules uniques dans des procédés de travail de génération d'évènements transgéniques et transplastomiques. L'invention élimine également les contraintes techniques et produit des lignées transgéniques s'exprimant sans marqueur et de manière uniforme avec un fort rendement pour supporter divers besoins de la santé animale, de la biopharmacie et de plateformes de protection de caractéristiques et de cultures.

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