TRANS-SPLICING TRANSCRIPTOME PROFILING

The present invention provides a method of identifying mRNA transcripts in the transcriptome of a cell comprising i) delivering into the cell a donor expression vector comprising nucleotides in a sequence encoding a trans-splicing barcode cassette, wherein the trans-splicing barcode cassette compris...

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Bibliographische Detailangaben
Hauptverfasser: ZNAMENSKIY PETR, PEIKON IAN D, ZADOR ANTHONY M
Format: Patent
Sprache:eng
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Zusammenfassung:The present invention provides a method of identifying mRNA transcripts in the transcriptome of a cell comprising i) delivering into the cell a donor expression vector comprising nucleotides in a sequence encoding a trans-splicing barcode cassette, wherein the trans-splicing barcode cassette comprises a) a first portion, the nucleotide sequence of which encodes an intron comprising as part of its 3′ end, or followed at its 3′ end by a splice-site nucleotide sequence; followed at its 3′ end by, b) a second portion, the nucleotide sequence of which encodes a barcode polynucleotide; followed at its 3′ end by c) a third portion, which encodes a nucleotide identification element sequence, ii) exposing the cell to conditions such that the cell produces multiple copies of the trans-splicing barcode cassette encoded by the donor expression vector, which multiple copies of the trans-splicing barcode cassette each splice the barcode polynucleotide onto a mRNA transcript of the cell, thereby forming multiple mRNA transcripts of the cell, each spliced to the barcode polynucleotide; and iii) identifying the multiple mRNA transcripts that are spliced to the barcode polynucleotides, thereby identifying mRNA transcripts in the transcriptome of the cell.