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Small RNAs that activate Lrig1 expression in glioma U251 cells, screening methods and their applications

A screening method, a technology for human glioma, applied in the field of double-stranded small activating RNA

Active Publication Date: 2020-06-23
CHINA THREE GORGES UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no effective treatment for glioma, making it a research focus in the field of neurosurgery

Method used

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  • Small RNAs that activate Lrig1 expression in glioma U251 cells, screening methods and their applications
  • Small RNAs that activate Lrig1 expression in glioma U251 cells, screening methods and their applications
  • Small RNAs that activate Lrig1 expression in glioma U251 cells, screening methods and their applications

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Embodiment 1

[0022] dsRNA Design dsLRIG1-712 is a dsRNA molecule with a length of 21 nucleotide base pairs designed for the relative transcription start point -712 of the LRIG1 gene promoter region and complementary to the promoter. The control dsRNA (dsControl) is non-homologous to the known human genome sequence, and is a dsRNA sequence with the same length of 21 nucleotide base pairs.

[0023] Using the Genebank database, find the sequence of the promoter region of the LRIG1 gene, and use the sequence upstream of the promoter -712 as a template for designing dsRNA. The specific sequence is: dsLRIG1-712: Sense: UUU CCC ACC CAA GGU GUG A[dT][dT]; Antisense: UCA CAC CUU GGG UGG GAA A[dT][dT].

[0024] Cell culture and transfection U251 cells were grown in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Incubator cultivation. The experiment was divided into 3 groups: blank group, control group (transfected with dsControl) and experimental group (transfected with candidate d...

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Abstract

The invention discloses a double-strand small activation RNA (dsLRIG-712) for activating cancer suppressor gene LRIG1 expression in brain glioma U251 cells and and application of double-strand small activation RNA (dsLRIG-712). dsLRIG-712 consists of 21 nucleotide positive-sense strands and antisense strands, two dTdT nucleotides are suspended at the 3'terminal of each strand in a protruding manner, and the other 19 nucleotides are mutually paired. The saRNA molecule can specifically activate expression of the LRIG1 gene, the target gene mRNA and the protein expression level are improved, proliferation and exercise capabilities of brain glioma cells are inhibited, and the purpose of suppressing tumor growth is achieved. Therefore, the saRNA molecule can be used for treating brain glioma.

Description

technical field [0001] The invention provides a double-stranded small activating RNA (dsLRIG-712) capable of activating the expression of tumor suppressor gene LRIG1 in glioma U251 cells, and uses dsLRIG-712 for the treatment of glioma. Background technique [0002] RNA activation (RNA activation, RNAa) is a phenomenon in which small molecule double-stranded RNA targets the gene promoter region and induces gene expression at the transcriptional level. The small dsRNA with activation function is called small activating RNA (small activating RNA, saRNA). As early as 1969, Britten et al. discovered that the RNA transcribed from the non-coding region of the genome can activate the expression of a large class of genes, and proposed the concept of activator RNAs (RNAa), but it was not confirmed. It was not until 2006 that saRNA was officially discovered and named by Li et al., who transfected prostate cancer PC-3 and DU- At 145, it was unexpectedly found that the target gene did...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A61K31/713A61P35/00G01N33/68
CPCA61K31/713C12N15/113C12N2310/10G01N33/68G01N2333/4704
Inventor 黄益玲胡火军孙雨沛李格尤程程
Owner CHINA THREE GORGES UNIV
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