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Knockout method of prion gene of goats

A prion and goat technology, applied in the field of biology, can solve problems such as the limitations of prion disease prevention and control research

Inactive Publication Date: 2012-10-03
YANGZHOU UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] Due to the particularity of the pathogenic factor Prion, people's research on the prevention and control of prion diseases is relatively limited

Method used

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  • Knockout method of prion gene of goats
  • Knockout method of prion gene of goats
  • Knockout method of prion gene of goats

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Embodiment Construction

[0030] 1. Characteristic materials: pSilencer 4.1-CMVneo vector (Ambion / Invitrogen # AM5779, siRNA-specific vector, used to load siRNA); pSG5 vector (Stratagene #216201, used to load the goat prion gene recognized by siRNA sequence for expression in mammalian cells); specific siRNA (sequence such as SEQ ID NO.1-6 ， Synthesized by Shanghai Invitrogen Company ） .

[0031] 2. According to the goat prion gene sequence (GenBank: X74758.1), use the program provided on the Ambion website to design three pairs of siRNA online as follows:

[0032] siRNA1: sense strand: AACCUGGCGGAGGAUGGAAtt (SEQ ID NO.1)

[0033] Antisense strand: UUCCAUCCUCCGCCAGGUUtt (SEQ ID NO.2)

[0034] siRNA2: sense strand: GCCACAUAGGCAGUUGGAUtt (SEQ ID NO.3)

[0035] Antisense strand: AUCCAACUGCCUAUGUGGCtt (SEQ ID NO.4)

[0036] siRNA3: sense strand: AAGCCACAUAGGCAGUUGGtt (SEQ ID NO.5)

[0037] Antisense strand: CCAACUGCCUAUGUGGCUUtt (SEQ ID NO.6);

[0038] The two single-stranded siRNAs of each pair of pri...

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Abstract

The invention belongs to the field of biology, and particularly relates to prion siRNA (small interfering ribonucleic acid) of goats and a knockout method of prion gene of goats. In the prion siRNA of goats, the sense strand sequence is shown as SEQ ID NO.1, while the antisense strand sequence is shown as SEQ ID NO.2. The knockout method of the prion gene of goats comprises guiding the prion siRNA of goats into a target cell through a vector for expressing so as to reduce the mRNA (messenger ribonucleic acid) level of the prion of goats. The method provides technical support to subsequent acquisition of an anti-prion transgenic goat breeding material.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a goat prion gene knockout method. Background technique [0002] Prion, also known as Prion protein, is a group of protein-soaked particles with a molecular weight of 27 000-30 000 Daltons, which can not find any nucleic acid so far, have strong resistance to various physical and chemical effects, and are infectious. Is the specific cause of transmissible spongiosa encephalopathy in humans and animals. Under certain conditions, the protein undergoes a mutation or configuration change, resulting in a change from benign to malignant, that is, to infectious disseminated particles, which can cause vacuolar degeneration of neurons in the brain of humans and animals (ie, spongy degeneration), so The disease caused by this type of virus is called transmissible spongiform encephalopathy (TSE), also known as prion disease. At present, there are about 30 kinds of zoonotic infectious di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12R1/93
Inventor 李拥军李太坤李文婷
Owner YANGZHOU UNIV
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