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Last Updated: January 29, 2022

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Claims for Patent: 7,149,631

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Summary for Patent: 7,149,631
Title:Alternative splicing and nonsense-mediated decay: computational methods and gene regulation
Abstract: Computational methods identify alternate splice forms of known gene transcripts and isoforms that are subject to NMD (nonsense-mediated decay). These methods were used to identify thousands of human genes that generate alternative splice forms, and to demonstrate that about a third of these are subject to NMD. This high prevalence of NMD-targeted transcripts indicates a systemic way of regulating gene expression--by shunting gene expression to nonproductive splice variants. This endemic regulation is exploited to engineer regulation of gene expression, to characterize splice pathway components and to assay splice environments, for example, using NMD-regulated reporter genes.
Inventor(s): Brenner; Steven E. (Oakland, CA), Green; Richard E. (Berkeley, CA), Lewis; Benjamin P. (Berkeley, CA)
Assignee: Regents of the University of California (Oakland, CA)
Application Number:10/159,997
Patent Claims:1. A computational method for systematically identifying alternative mRNA splice isoforms of known genes, the method comprising the steps of: a) identifying target gene sequences by mapping mRNA sequences of an mRNA sequence dataset to genomic sequences of a genomic DNA sequence dataset, wherein: each mRNA sequence is required to align to a corresponding genomic sequence over the full length of the coding sequence of the mRNA sequence, without gaps in the exons of the genomic sequence; and at least 98% identity between each mRNA sequence and the corresponding genomic sequence is required, favoring the mRNA sequence in case of nucleotide mismatch; and b) identifying a dataset of alternate mRNA splice isoforms of the target gene sequences by aligning EST sequences from an EST sequence dataset to the target gene sequences and using a transcript assembly protocol, wherein: the alternative mRNA splice isoform dataset is restricted to mRNA sequences in which the 5' end of an EST sequence aligns to a coding sequence of the corresponding mRNA sequence, such that the reading frame of the coding sequences can be determined for all isoforms of the dataset, and isoforms presenting intron retention are excluded from the alternative mRNA splice isoform dataset, and coverage by multiple EST sequences is required for each splicing event.

2. A computational method according to claim 1, further comprising the step of identifying a subset of the alternative mRNA splice isoform dataset consisting of isoforms subject to nonsense-mediated decay (NMD), comprising: screening the alternative mRNA splice isoform dataset for a subset of isoforms comprising alternate splices which introduce a stop codon more than 50 bp upstream of the final exon-exon splice junction, and classifying such subset isoforms as comprising premature stop codons and as targets of NMD.

3. A computational method according to claim 1, further comprising the step of correlating the alternatively spliced isoforms with their relative expression in cancer cells and non-cancer cells.

4. A computational method according to claim 2, further comprising the step of correlating the alternatively spliced isoforms with their relative expression in cancer cells and non-cancer cells.

Details for Patent 7,149,631

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 1986-06-04 ⤷  Try it Free 2039-02-26
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try it Free 2039-02-26
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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