SiRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression and application thereof

A gene expression, p21 technology, applied in the field of siRNA

Active Publication Date: 2014-04-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for specific and efficient silencing of genes involved with inflammatory processes such as lipids or fats from blood vessels caused by excessive alcohols (ethanol). It also helps researchers identify potential causes related diseases like Non Alcoholics Fat Liver Disease (NAFLD) through studying these changes over time.

Problems solved by technology

Technologies aim at studying how nonspecific small RNAs called miR-39 play key roles during the course of developing new treatments for inflammatory bowel disease (IBD), including those associated with necrosis and autoimmunity. Previous methods were limited in predictive ability, making them challenging to apply these techniques effectively without causing complications like acute renal injury.

Method used

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  • SiRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression and application thereof
  • SiRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression and application thereof
  • SiRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, siRNA design and synthesis

[0023] 1. Synthesis of p21 siRNA

[0024] The full-length human P21 mRNA sequence (NM_000389.4; SEQ ID NO.5) was obtained from the National Center for Biotechnology Information (NCBI) database, and according to the principle of RNAi, combined with design software, the design, synthesis and An effective 21nt siRNA against human p21 gene was screened out for four transcripts. A BLAST comparison check was performed to ensure that there was no homology to other genes.

[0025] The p21 siRNA sequence is:

[0026] 5'-GUCACUGUCUUGUACCCUUTT-3' (sense strand) (SEQ ID NO.1); 5'-AAGGGUACAAGACAGUGACTT-3' (antisense strand) (SEQ ID NO.2).

[0027] Chemical synthesis by Shanghai Gemma Pharmaceutical Technology Co., Ltd. (factory address: 602, No. 1011 Halei Road, Zhangjiang, Pudong, postcode: 201203): using NTP as raw material, using ABI3900 nucleic acid synthesizer to chemically synthesize single-stranded RNA, and finally in the annealin...

Embodiment 2

[0033] Example 2, the effect of p21 siRNA on the expression of p21 gene in L02 cells in vitro

[0034] 1. Group transfection

[0035] Spread L02 cells evenly in a 6-well plate and divide them into 4 groups:

[0036] High-fat experiment group: DMEM / F12 medium containing 10% (volume percentage) inactivated neonatal bovine serum, 666 μmol / L sodium oleate and 333 μmol / L sodium palmitate; transfection 200 pmol Example 1 p21 siRNA.

[0037] High-fat control group: DMEM / F12 medium containing 10% (volume percentage) inactivated neonatal bovine serum, 666 μmol / L sodium oleate and 333 μmol / L sodium palmitate; transfection 200 pmol Example 1 negative control siRNA.

[0038] Normal experimental group: DMEM / F12 medium containing 10% (volume percent) inactivated neonatal bovine serum, 100 U / ml penicillin and 100 mg / ml streptomycin; transfected with 200 pmol p21 siRNA of Example 1.

[0039] Normal control group: DMEM / F12 medium containing 10% (volume percentage) inactivated neonatal bovi...

Embodiment 3

[0044] Example 3, Effect of ECHS1 siRNA on L02 Cell Steatosis in Vitro

[0045] 1. Group transfection

[0046] The L02 cells were spread evenly in a 6-well plate and divided into 4 groups. Concrete grouping is with the step one of embodiment 2.

[0047] 2. The effect of p21 siRNA on lipid change of L02 cells in vitro

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Abstract

The invention discloses a siRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression and an application of the siRNA. The siRNA for specifically inhibiting p21 gene expression is a double-stranded RNA formed by nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2. The modified RNA is obtained by modifying the double-stranded RNA. In vitro test results show that the RNA provided by the invention can specifically and efficiently inhibit p21 gene expression so as to reduce lipid droplets and inhibit deposition of triglyceride. The siRNA provided by the invention can be applied to researching pathogenesis of non-alcoholic fatty liver diseases and potential researching non-alcoholic fatty liver diseases. The siRNA (Ribonucleic Acid) for specifically inhibiting p21 gene expression has an important value.

Description

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Claims

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Application Information

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Owner ZHEJIANG UNIV
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