Screening methods in eucaryotic cells

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12N 1/14 (2006.01) C12N 5/00 (2006.01) C12N 15/00 (2006.01) C12N 15/09 (2006.01) C12N 15/10 (2006.01) C12P 21/04 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2285435

The invention provides new methods for screening libraries of peptides and other compounds for a desired property in eucaryotic cells. The methods are premised, in part, on the unexpected observation that the contents of procaryotic or lower eucaryotic cells, such as yeast, can be transferred to recipient eucaryotic cells in an essentially clonal manner by protoplast fusion of the respective cells. Applications of the methods include screening peptides in eucaryotic cells substantially incapable of episomal replication of transferred nucleic acid fragments; screening in eucaryotic cells peptides or secondary metabolites produced in procaryotic cells; and screening a library of peptides for capacity to bind a selected RNA in eucaryotic cells.

L'invention concerne de nouvelles méthodes de criblage de banques de peptides et d'autres composés pour obtenir une propriété voulue dans des cellules eucaryotes. Les méthodes sont en partie fondées sur l'observation inattendue que le contenu de cellules procaryotes ou eucaryotes inférieures, telles que la levure, peut être transféré dans des cellules eucaryotes réceptrices d'une manière essentiellement clonale par fusion de protoplastes des cellules respectives. Des applications des méthodes selon l'invention comprennent le criblage de peptides dans des cellules eucaryotes essentiellement incapables d'une réplication épisomique de fragments d'acide nucléique transférés; le criblage dans des peptides de cellules eucaryotes ou des métabolites secondaires produits dans des cellules procaryotes; et le criblage d'une banque de peptides pour leur capacité à se lier à un ARN sélectionné dans des cellules eucaryotes.

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