Test indicator based on a single cell to control gene expression patterns with high spatial and temporal resolution

摘要

Procedure in vitro study of gene transcription in a cell or in a cell line comprendelas following steps: a. providing a cell or a cell line, excluding a cell or human cell line embryonic whichthe was introduced a double stranded polynucleotide comprising said positive strand considered from its 5 'end to its 3' end sobresu double-stranded polynucleotide, (i) a promoter of a gene of interest several promoters of various genes of interest selected among genes which are endogenous to the cell, and are subjected to a profiling of gene expression wherein said promoter is recognized by elmecanismo inner transcription of the cell, and (ii) one or more barcodes beacon in quecada barcode contains at least one unit of barcode that is a DNA quecomprende tandem repeats of at least one binding site recognition mark , estandocada binding site composed of a nucleotide sequence, and wherein each of said codes is under debarras controlling at least one of said promoters for transcription; b. provoke silencing or modulating transcription of the polynucleotide assembly, c. contacting the cell or cell line of step a with the probe (s) (s) detection can hibridarcon (the) site (s) recognition binding beacon of (the) code (s) bars and are a ovarias molecular beacons that can hybridise with (the) site (s) binding beacon (of) code (s) debarras, said (s) beacon (s) molecular structure polynucleotide stem-loop and resultandoaptas for visualization or measurement when they are hybridized to their target sequence, especially reversible unamanera, in particular wherein the display of the hybridization probe (s) (s) consu detection target is obtained as result of fluorescence which is activated when the detection probe sequence she joined the Dutchpro diana.d. detecting hybridization between the detection probes and the transcription of derecognition binding sites of the barcode as an indicator of transcription activity of the promoter delmontaje polynucleotide.