Single cell based reporter assay to monitor gene expression...

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01)

Patent

CA 2699358

The invention relates to a double-stranded polynucleotide comprising on its positive strand considered from its 5' end to its 3' end, (i) a promoter of a gene of interest or several promoters of various genes of interest selected among genes which are endogenous to a determined cell, and, (ii) one or several barcode(s) wherein each barcode contains at least one barcode unit formed of at least one, especially of multiple, recognition binding sites each binding site being composed of a nucleotide sequence, and wherein each of said barcode(s) is under the control of at least one of said promoter(s) for transcription. It further concerns use of said polynucleotide to monitor gene expression patterns in living cells, especially in single cells, with a rapid and high spatio-temporal resolution.

La présente invention concerne un polynucléotide double brin comprenant sur son brin positif pris de son extrémité 5' à son extrémité 3', (i) un promoteur d'un gène intéressant ou plusieurs promoteurs de divers gènes intéressants choisis parmi les gènes qui sont endogènes à une cellule déterminée, et, (ii) un ou plusieurs codes à barres, chacun desdits codes contenant au moins une unité de code à barres formée d'au moins un, et tout particulièrement de multiples, sites de liaison de reconnaissance, chaque site de liaison étant composé d'une séquence nucléotidique, et chacun desdits codes à barres étant sous le contrôle d'au moins un desdits promoteurs pour la transcription. L'invention concerne en outre l'utilisation dudit polynucléotide pour contrôler des profils d'expression génique dans des cellules vivantes, notamment dans des cellules individuelles, avec une résolution spatio-temporelle rapide et élevée.

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