High efficiency siRNA molecule to target gene Rig-G

A gene and purpose technology, applied in the field of small double-stranded RNA molecules, can solve the problems of inhibiting AP-1 protein, affecting binding, and reducing stability.

Inactive Publication Date: 2006-08-02
RUIJIN HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

We think that the change in the distribution of JAB1 protein in the cell may be caused by the interaction of Rig-G protein located in the cytoplasm and the interaction with JAB1 to retain part of JAB1 in the cytoplasm, and then affect the combination of JAB1 and c-Jun , resulting in a decrease in the stability of c-Jun binding to its corresponding response element, thereby inhibiting the transcriptional activity of the AP-1 protein

Method used

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  • High efficiency siRNA molecule to target gene Rig-G
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  • High efficiency siRNA molecule to target gene Rig-G

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] High-efficiency siRNA to inhibit expression of target gene Rig-G (1)

[0031] 1. Select the sequence on the target gene and synthesize the DNA template:

[0032] According to the basic characteristics of effective RNAi sequences, we selected sites in the cDNA sequence of the target gene Rig-G gene (see Table 1 and Table 2) and synthesized DNA templates (synthesized by Boya Biotech).

[0033] Basic requirements for selecting candidate siRNA sequences on target genes:

[0034] 1) Search for a candidate sequence (but not absolute) at a position 50-100 bp downstream of the promoter of the coding region of the target gene.

[0035] 2) The two template strands (forward strand and anti-strand) of the siRNA duplex: both start with AA to ensure that the 3' of the siRNA duplex transcribed in vitro protrudes two UUs.

[0036] 3) Among the paired 19 nucleotides, the first position is G, and the nineteenth position is C.

[0037] 4) The GC content is preferably 40-55%.

[0038] ...

Embodiment 2

[0061] High-efficiency siRNA to inhibit the expression of target gene Rig-G (2)

[0062] 1. Select the sequence on the target gene and synthesize the DNA template:

[0063] According to the basic characteristics of effective RNAi sequences, we selected sites in the cDNA sequence of the target gene Rig-G gene (see Table 1 and Table 2) and synthesized DNA templates (synthesized by Boya Biotech).

[0064] Basic requirements for selecting candidate siRNA sequences on target genes:

[0065] 1) Search for a candidate sequence (but not absolute) at a position 50-100 bp downstream of the promoter of the coding region of the target gene.

[0066] 2) The two template strands (forward strand and anti-strand) of the siRNA duplex: both start with AA to ensure that the 3' of the siRNA duplex transcribed in vitro protrudes two UUs.

[0067] 3) Among the paired 19 nucleotides, the first position is G, and the nineteenth position is C.

[0068] 4) The GC content is preferably 40-55%.

[00...

Embodiment 3

[0090] Efficient siRNA to inhibit expression of target gene Rig-G (3)

[0091] 1. Select the sequence on the target gene and synthesize the DNA template:

[0092] According to the basic characteristics of effective RNAi sequences, we selected sites in the cDNA sequence of the target gene Rig-G gene (see Table 1 and Table 2) and synthesized DNA templates (synthesized by Boya Biotech).

[0093] Basic requirements for selecting candidate siRNA sequences on target genes:

[0094] 1) Search for a candidate sequence (but not absolute) at a position 50-100 bp downstream of the promoter of the coding region of the target gene.

[0095] 2) The two template strands (forward strand and anti-strand) of the siRNA duplex: both start with AA to ensure that the 3' of the siRNA duplex transcribed in vitro protrudes two UUs.

[0096] 3) Among the paired 19 nucleotides, the first position is G, and the nineteenth position is C.

[0097] 4) The GC content is preferably 40-55%.

[0098] 5) Homol...

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Abstract

The present invention relates to one kind of small segment double stranded RNA molecule (siRNA) with high efficiency interference on the expression of target gene Rig-G. The high efficiency siRNA is prepared through the following steps: 1. selecting site in the cDNA sequence of target gene to synthesize DNA template; 2. extracorporeal transcription to synthesize RNA segment; or constructing siRNA expression plasmid. Using these siRNA can inhibit the expression of Rig-G gene effectively and lay foundation for establishing Rig-G deficiency type cell strain mold, and is favorable to understanding the effect of Rig-G in vitamin A acid signal conduction network and revealing the molecular mechanism for vitamin A acid to induce APL cell differentiation.

Description

technical field [0001] The invention relates to the fields of molecular genetics, cell biology and hematology. Specifically, the present invention relates to a class of small-fragment double-stranded RNA molecules capable of efficiently interfering with the expression of the target gene Rig-G. Background technique [0002] Acute promyelocytic leukemia (APL) is a more aggressive subtype of acute myeloid leukemia. my country's medical community took the lead in the world to use all-trans retinoic acid (ATRA) differentiation induction therapy for APL successfully, which became a new milestone in the history of tumor treatment. In order to elucidate the mechanism of ATRA-induced differentiation of APL cells, scholars from various countries have conducted a lot of research for more than ten years. But so far the signaling pathway of retinoic acid is still a mystery. In order to make a breakthrough in this research, Shanghai Institute of Hematology has been working on the resea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34C07H21/02C12N15/63
Inventor 童建华肖澍贾培敏
Owner RUIJIN HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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