In-vivo gene silencing method taking lentivirus as tool

A technology of gene silencing and lentivirus, which is applied in the fields of molecular genetic biology and developmental biology, can solve the problems of high cost, side effects, and time-consuming, and achieve the effect of simple method

Active Publication Date: 2017-07-07
QINGDAO UNIV
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Problems solved by technology

[0005] To sum up, the problems existing in the existing technology are: the existing gene manipulation methods in whole organisms are costly and time-consuming, and are likely to cause unknown side effects, while the lentivirus-mediated siRNA silencing basi

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  • In-vivo gene silencing method taking lentivirus as tool

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Embodiment Construction

[0019] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0020] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0021] Such as figure 1 As shown, the in vivo gene silencing method using lentivirus as a tool provided by the embodiment of the present invention includes the following steps:

[0022] S101: Use a 5ul micro-injector to inject the virus, the injection volume is set to 1ul per 20g egg weight, and the virus titer is uniformly 3x10 8 TU / ml;

[0023] S102: 2-day-old chicken embryos, open a hole with a diameter of about 1.5-2 mm in the air cell, visually determine the position of the air...

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Abstract

The invention belongs to the field of molecular genetic biology and development biology and discloses an in-vivo gene silencing method taking lentivirus as a tool. The in-vivo gene silencing method taking the lentivirus as the tool comprises the following steps: injecting virus by utilizing a microsyringe; after 2 days, drilling a hole in an air chamber of a chicken embryo and watching and determining the position of an air chamber membrane; inserting the microsyringe into a median membrane of the air chamber to inject the virus. The invention establishes the in-vivo gene silencing method taking the lentivirus as the tool and also successfully establishes a PPARalpha in-vivo silencing chicken embryo model. The method provided by the invention is relatively simple and convenient and operation can be finished under a visual condition; the method is low in cost and high in practicability and the survival rate can reach 80 percent; the transfection efficiency on a heart is high and the in-vivo silencing efficiency can reach about 70 percent. A complicated development signal transduction process is completely kept by in-vivo transfection so that the method can be used as a powerful tool for researching effects of a target gene in development.

Description

technical field [0001] The invention belongs to the fields of molecular genetic biology and developmental biology, and in particular relates to an in vivo gene silencing method using lentivirus as a tool. Background technique [0002] Gene manipulation is an essential method to study the role of a specific gene in life activities. The classic method is gene knockout (Knock out), which can effectively remove the target gene, but it is relatively expensive and time-consuming. At the same time, due to its Changes to the biological genome may bring some unknown side effects and reduce the reliability of the research. [0003] siRNA is a small molecule RNA discovered in recent years that can specifically recognize, inhibit and degrade specific mRNA, thereby reducing the protein expression level of the target gene without changing the DNA. It has been widely used to silence the target gene to study its role in specific biological activities. Common methods for introducing siRNA ...

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Application Information

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IPC IPC(8): C12N15/89C12N5/10
CPCC07K14/465C12N5/0603C12N15/89
Inventor 姜启晓赵萌韩彦弢
Owner QINGDAO UNIV
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