Compositions and methods for specific cleavage of exogenous rna in a cell

A composition and exogenous technology, applied in the field of compositions for polynucleotide expression, can solve the problems of unsuitable cancer-derived signal sequences, harmful side effects and the like

Inactive Publication Date: 2013-09-04
NANODOC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this approach has several inherent problems: the first is that the RNA molecule containing the signal sequence must be present in the cell in very high copy numbers because trans-splicing events are very rare
A second problem is that in most cases the method is not suitable for signal sequences of cancer origin, where mutations spread over short regions
A third problem is that trans-splicing events can also occur randomly and thus can lead to unwanted side effects
A fourth problem is that RNA molecules containing signaling molecules must contain an intron at a very specific location
Another method that won the 2004 World Technology Award in Biotechnology proposed the use of small dsDNA, ssDNA, hairpin DNA and restriction enzymes, however this method can only be extracted from cells under very unique conditions function in organisms rather than under the physiological conditions of living cells [20]

Method used

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  • Compositions and methods for specific cleavage of exogenous rna in a cell
  • Compositions and methods for specific cleavage of exogenous rna in a cell
  • Compositions and methods for specific cleavage of exogenous rna in a cell

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Embodiment approach

[0436] This section defines and describes additional embodiments of the compositions of the invention described in any of the preceding embodiments in any of the preceding sections.

[0437]Endogenous signal RNA may be, for example, but not limited to: viral RNA comprising a predetermined signal sequence, cellular RNA such as, for example, mRNA and the like. The predetermined signal sequence can be, for example, a signal sequence unique to neoplastic cells, a signal sequence of viral origin, and the like, or a combination thereof. In certain embodiments, the predetermined signal sequence does not comprise any other type of endogenous RNA molecule capable of directing or effecting cleavage of the RNA molecule within the cell (such as, for example, miRNA, shRNA, ribozyme, stRNA, and similar RNA molecules) .

[0438] According to certain embodiments, cells that may be used in embodiments of the present invention may be any type of cell of any origin, such as, for example, but no...

Embodiment 1

[0496] Example 1 : Use of the composition of the present invention to kill cancer cells in a specific patient

[0497] According to the American Cancer Society, during 2007, 7.6 million people worldwide died of cancer.

[0498] In this example the compositions of the invention are designed to specifically kill cancer cells in a particular patient. The first step in designing a composition of the invention for a particular patient is to identify a predetermined signal sequence, which is an 18-25 nucleotide long sequence of an endogenous RNA molecule present in cancer cells of that particular patient, wherein the predetermined signal sequence is absent from any endogenous RNA molecule in healthy or non-metastatic tumorigenic cells of the body of that particular patient. Thus, the predetermined signal sequence is the RNA sequence of a gene mutated in cancer cells. On average each tumor contains mutations in 90 protein-coding genes [16] and each tumor originates from a single ...

Embodiment 2

[0509] Example 2 : Use of the composition of the present invention to kill EBV-related gastric cancer cells, nasopharyngeal cancer cells, Burkitt's lymphoma cells and Hodgkin's lymphoma cells

[0510] Epstein-Barr virus (EBV) is a ubiquitous human gammaherpesvirus that establishes a lifelong latent infection in B lymphocytes after initial infection. EBV infects most of the human population worldwide and has been implicated in the pathogenesis of several human malignancies, including Burkitt and Hodgkin lymphomas, gastric cancer, and nasopharyngeal carcinoma (NPC) [32]. A major feature of EBV infection is the expression of late genes including EBNA1, LMP1, LMP2, and EBER [32]. LMP1 (latent membrane protein 1) was the first EBV latent gene discovered to be able to transform cell lines and alter cellular phenotype due to its oncogenic potential [32]. In human epithelial cells, LMP1 alters many functional properties involved in tumor progression and invasion [32].

[0511] In ...

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Abstract

There are provided compositions for cleaving an exogenous RNA of interest only in the presence of an endogenous signal RNA sequence, thereby activating expression of a polynucleotide of interest only in the presence of the endogenous signal RNA sequence. There are provided methods for the preparation of the composition and uses thereof in treatment and diagnosis of various conditions and disorders, for example by selectively activating expression of a toxin only in specific target cell populations.

Description

field of invention [0001] The present invention relates to compositions for cleaving an exogenous RNA of interest only in the presence of an endogenous signal RNA sequence, thereby activating the expression of a polynucleotide of interest only in the presence of an endogenous signal RNA sequence. The present invention also relates to the use of these compositions in the treatment and diagnosis of various conditions and disorders, as exemplified by selectively activating the expression of the toxin only in target cell populations. Background of the invention [0002] RNA interference (RNAi) is a phenomenon in which dsRNA consisting of sense RNA and antisense RNA homologous to a specific region of a target gene achieves cleavage of the homologous region of the transcript of the target gene, thereby inhibiting gene expression. In mammals, dsRNA should be shorter than 31 base pairs to avoid triggering an interferon response that can lead to cell death through apoptosis. RNAi te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/00
CPCA61P31/12A61P35/00C07H21/04C12N15/113C12N2310/14C12N15/63
Inventor 盖伊·阿维特沃尔
Owner NANODOC
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