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ShRNA capable of reducing cell apoptosis caused by fluorine and application of shRNA

A cell and apoptosis technology, applied in shRNA and its application field, can solve interference and other problems, and achieve the effect of reducing fluorine-induced apoptosis

Active Publication Date: 2018-04-03
GUIZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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However, there is no effective means of interfering

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  • ShRNA capable of reducing cell apoptosis caused by fluorine and application of shRNA
  • ShRNA capable of reducing cell apoptosis caused by fluorine and application of shRNA
  • ShRNA capable of reducing cell apoptosis caused by fluorine and application of shRNA

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specific Embodiment approach

[0032] The present invention provides a kind of shRNA that can reduce fluorine-induced apoptosis, and the specific implementation method is as follows:

[0033] (1) Design and screening of PRKAR1A gene siRNA

[0034] 1) Design of PRKAR1A gene siRNA

[0035] According to the comparison results of the PRKAR1A whole gene mRNA of rat (NM.013181.1) and mouse (NM.021880.2) in NCBI, the two have high homology, so a common siRNA sequence for rat and mouse was designed. BLAST (NCBI) is used for non-homologous query of the sequence. The siRNA target has no other homologous sequences. The siRNA is designed to target the ORF region (open reading frame region) of the target gene. By comparing the same sequence, a 21-base positive-sense strand and The complementary base antisense strands constitute three template strands in total, as shown in Table 1.

[0036] Table 1 siRNA sequence

[0037]

[0038] 2) Screening of PRKAR1A gene siRNA

[0039] The 3 pairs of synthetic siRNAs were tra...

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Abstract

The invention discloses a shRNA capable of reducing cell apoptosis caused by fluorine and application of the shRNA. The shRNA comprises a sense strand nucleotide and an antisense strand nucleotide, the sequence of the sense strand nucleotide is shown in SEQ NO.1, and the sequence of the antisense strand nucleotide is shown in SEQ NO.2. A PRKAR1A shRNA interfered UMR-106 and MC3T3-E1 cell model isexposed to fluorine for 48 hours through the shRNA technique, the expression of proteins of Cleaved Caspase-3, Caspase-8, Bim and PARP is detected through Western Blot, the cell apoptosis is detectedthrough flow cytometry and TUNREL method, thereby proving that PRKAR1A shRNA plays a certain role in the fluorine-induced osteoblast apoptosis, and a shRNA fragment can reduce the cell apoptosis caused by fluorine.

Description

technical field [0001] The invention belongs to the technical field of bioengineering gene recombination, and in particular relates to a shRNA capable of reducing fluorine-induced apoptosis and its application. Background technique [0002] As a chemically active non-metallic trace element necessary for the body, fluorine is widely used in production and life. Adequate fluoride is of great significance for the prevention of dental caries and the treatment of osteoporosis, and long-term excessive intake will cause a chronic systemic disease characterized by skeletal fluorosis and dental fluorosis. Disease, namely endemic fluorosis (endemic fluorosis). As one of the main target organs of fluoride, bone is composed of osteoblasts and osteoclasts. Osteoblasts are an important part of bone. The balance between bone formation caused by osteoblasts and bone resorption caused by osteoclasts is the key to maintaining bone integrity. When the relationship between them is out of bala...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/861A61P39/02
CPCC12N15/113C12N15/86C12N2310/14C12N2710/10043
Inventor 韦艳
Owner GUIZHOU MEDICAL UNIV