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Natural antisense and non-coding RNA transcripts as drug targets

A technology of antisense transcripts, transcripts, applied in the field of natural antisense and non-coding RNA transcripts as drug targets

Active Publication Date: 2009-05-20
THE SCRIPPS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And, until a few years ago, there was no systematic effort to identify novel ncRNA transcripts and elucidate their functions

Method used

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  • Natural antisense and non-coding RNA transcripts as drug targets
  • Natural antisense and non-coding RNA transcripts as drug targets
  • Natural antisense and non-coding RNA transcripts as drug targets

Examples

Experimental program
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preparation example Construction

[0207] The preparation method of the hybridization probe specific to the DNA encoding the target gene comprises: cloning the oligonucleotide sequence encoding the target gene or its derivatives into the vector used for preparing the mRNA probe. Such vectors are well known to those skilled in the art, are commercially available, and can be used to synthesize RNA probes in vitro by adding appropriate RNA polymerase and appropriate labeled nucleotides. Hybridization probes can be probed with various reporter groups (e.g., radionuclides such as 32 P or 32 S) or an enzyme label (such as alkaline phosphatase coupled to the probe through an avidin-biotin coupling system, a fluorescent label, etc.). The polynucleotide sequence encoding the target gene can be used for: Southern analysis (Southern blot) or Northern analysis (Northern blot), dot blot, or other membrane-based techniques; PCR techniques; dipsticks, pins, and multiple ELISA-like assays ; Using microarrays of fluid and tis...

Embodiment 1

[0300] Example 1: Knockdown of Antisense Transcripts

[0301] In the context of the present invention, the only way to affect the levels of antisense transcripts is to use siRNAs designed to efficiently knock down transcripts. The later term - knockdown - was introduced when we were studying antisense oligonucleotides in the early 1990s (Wahlestedt, C. (1994) Antisense oligonucleotide strategies in neuropharmacology. Trends Pharmacol Sci 15(2): 42- 46), but the same applies to siRNA.

[0302] Expression profiles show frequent consistent regulation of sense / antisense pairs. Using siRNA, we provide experimental evidence that scrambling of antisense RNA by siRNA can alter the expression of the corresponding sense messenger RNA. However, this regulation can be discordant (antisense knockdown causes an increase in the sense transcript) or consistent (antisense knockdown causes a concomitant decrease in the sense transcript). Some human and mouse antisense transcripts that have b...

Embodiment 2

[0314] Example 2: Natural antisense-mediated regulation of gene expression in mammals

[0315] Naturally occurring antisense transcripts (NATs) have been reported for 20% of the human genome. A recent report indicated that at least 72% of murine transcripts present NAT. Most natural antisense transcripts are cis-encoded antisense. Cis-NAT is defined as complementary mRNAs with overlapping transcription units at the same chromosomal locus. Trans-NAT is a complementary mRNA transcribed from a different chromosomal location. Chimeric transcripts are mRNAs that share identity to more than one region of the genome, and can be artifacts of cDNA library products. More than 70% of cis-NATs had a tail-to-tail pattern of 3' overlap, while 15% had a head-to-head pattern of 5' overlap. The remaining molecules have overlaps of ingenic regions or coding sequences. Many NATs do not exhibit open reading frames and are classified as non-coding RNAs.

[0316] Interactions between antisens...

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PUM

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Abstract

Small interfering RNA (siRNA) knock down antisense transcripts, and regulate the expression of their sense partners. This regulation can either be discordant (antisense knockdown results in sense transcript elevation) or concordant (antisense knockdown results in concomitant sense transcript reduction).

Description

technical field [0001] The present invention provides compositions and methods for highly selective targeting of heterologous nucleotide sequences. Oligonucleotides are siRNA's that bind to their target genes in a sequence-dependent manner and are capable of modulating the expression of non-essential nucleotide sequences in target cells. Moreover, siRNA's can specifically and selectively kill bacteria or human cells if the target is present in the genome of the bacteria or human cells. Background technique [0002] Our understanding of the complexity of mammalian transcription has improved dramatically over the past few years, and multiple novel ribonucleic acid (RNA) transcripts have been detected. This was somewhat of a surprise, since the total number of conventional (protein-coding) genes in the human genome (approximately 20,000-25,000) is much lower than expected in previous years, and compared with the simpler organisms Drosophila melanogaster or The gene numbers of...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12Q1/68C07H21/04
CPCC12N15/113C12N2310/14C12N2330/10C12N2310/111A61P1/04A61P3/10A61P5/14A61P7/08A61P9/00A61P9/10A61P11/00A61P13/12A61P17/00A61P17/02A61P17/06A61P19/02A61P21/00A61P25/00A61P25/02A61P25/04A61P25/08A61P25/14A61P25/16A61P25/18A61P25/20A61P25/22A61P25/24A61P25/28A61P25/32A61P25/36A61P27/02A61P27/16A61P29/00A61P31/00A61P31/04A61P35/00A61P35/02A61P37/02A61P37/06A61P37/08A61P39/02A61P43/00
Inventor C·瓦勒斯泰特
Owner THE SCRIPPS RES INST
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