Populations of reporter sequences and methods of their use

C - Chemistry – Metallurgy – 40 – B

Patent

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Details

C40B 40/06 (2006.01) A01K 67/027 (2006.01) C07H 21/00 (2006.01) C12N 5/10 (2006.01) C12N 15/65 (2006.01) C12Q 1/00 (2006.01) C12Q 1/04 (2006.01) C12Q 1/68 (2006.01) C40B 40/02 (2006.01) G01N 33/68 (2006.01) G06F 19/00 (2006.01)

Patent

CA 2587338

Compositions, methods and kits are provided that are useful, for example, for determining activities of multiple cis-regulatory sequences, such as promoters and enhancers, and/or multiple trans-acting factors, such as transcription factors, in a cell. In particular, in certain embodiments, compositions are provided comprising a population of polynucleotide reporter transcription units (RTUs) in which each RTU comprises a reporter sequence, a processing tag located in the reporter sequence; and a cis-regulatory element operably linked to the reporter sequence, wherein the reporter sequences between any two RTUs in the population, outside of the processing tags, are substantially identical and wherein the positions of the processing tags within the reporter sequences distinguish between any two RTUs differing, for example, in their cis- regulatory elements. The compositions, methods and kits can further be used, for example, to identify a cell type or disease state, for example, in a biological organism.

L'invention concerne des compositions, des procédés et des trousses utiles, par exemple, pour déterminer dans une cellule les activités de multiples séquences régulatrices en cis, telles que des promoteurs et des amplificateurs, et/ou de multiples facteurs actifs en trans, tels que des facteurs de transcription. En particulier, certaines formes de réalisation se réfèrent à des compositions comprenant une population d'unités de transcription de rapporteur polynucléotidique (RTU), chaque RTU comprenant: une séquence rapporteur, une étiquette de traitement placée dans la séquence rapporteur et un élément régulateur en cis, lié de façon opérationnelle à la séquence rapporteur. Les séquences rapporteur qui se situent entre deux RTU de la population, quelles qu'elles soient, à l'extérieur des étiquettes de traitement, sont sensiblement identiques; et la position des étiquettes de traitement à l'intérieur des séquences rapporteur permet de distinguer deux RTU dont les éléments régulateurs en cis, par exemple, sont différents. Les compositions, procédés et trousses peuvent en outre servir, par exemple, à identifier un type de cellule ou un état pathologique, par exemple dans un organisme biologique.

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