Monkey encephalic zone virus injection method and applications thereof

A technology of viruses and brain regions, which is applied to the virus injection method and application field of monkey brain regions, can solve the problems of difficult virus diffusion, consumption of large viruses, and high virus titer requirements, and achieve the goal of improving expression efficiency and efficiency Effect

Inactive Publication Date: 2018-12-07
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, using the above method, the effect of virus expression in the brain area of ​​monkeys often does not meet the experimental requirements
Among them, the primary technical difficulty is that the virus itself will cause the immune response of the monkey body to produce antibodies to neutralize the virus
Secondly, it is difficult for the virus to spread to a large area when it is only injected from one site; injecting a larger area than the brain requires a large amount of virus consumption, and the

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Injection of virus into the brain area of ​​monkeys

[0042] (1) Virus packaging: use DNA ligase to connect the plasmid vector fragment carrying the GFP gene with complementary cohesive ends and the modified adenovirus genome to construct a viral plasmid; transfect the viral plasmid into 239T cells to carry out the viral plasmid Proliferation; then purification, titer test and expression test are carried out to obtain the packaged virus;

[0043] (2) Virus modification: chemically couple magnetic induction protein, motor protein and ATP on the exposed surface residues of the packaged virus shell to form a modified virus;

[0044] (3) Virus injection: After the monkey is anesthetized, the spinal cord is peeled off with a scalpel, and a hole is formed on the spinal cord with a skull drill to form a wound, and the modified virus is injected into the spinal fluid from the wound;

[0045] (4) Locating the target brain area: first glue the wound with biological tis...

Embodiment 2

[0048] Example 2 Injection of virus into the brain region of monkeys

[0049] (1) Virus packaging: use DNA ligase to connect the plasmid vector fragment carrying the GFP gene with complementary cohesive ends and the modified adenovirus genome to construct a viral plasmid; transfect the viral plasmid into 239T cells to carry out the viral plasmid Proliferation; then purification, titer test and expression test are carried out to obtain the packaged virus;

[0050] (2) Virus modification: chemically couple magnetic induction protein, motor protein and ATP on the exposed surface residues of the packaged virus shell to form a modified virus;

[0051] (3) Virus injection: After the monkeys were anesthetized, the spinal cord was peeled off with a scalpel, and a wound was formed on the spinal cord with a skull drill, and the modified virus was injected into the spinal fluid from the wound; Embodiment 1 is identical;

[0052] (4) Locating the target brain area: first glue the wound ...

Embodiment 3

[0055] Example 3 Injection of virus into the brain region of monkeys

[0056] (1) Virus packaging: use DNA ligase to connect the plasmid vector fragment carrying the GFP gene with complementary cohesive ends and the modified lentiviral genome to construct a viral plasmid; transfect the viral plasmid into 239T cells for viral plasmid Proliferation; then purification, titer test and expression test are carried out to obtain the packaged virus;

[0057] (2) Virus modification: chemically couple magnetic induction protein, motor protein and ATP on the exposed surface residues of the packaged virus shell to form a modified virus;

[0058] (3) Virus injection: After the monkeys were anesthetized, the spinal cord was peeled off with a scalpel, and a wound was formed on the spinal cord with a skull drill, and the modified virus was injected into the spinal fluid from the wound; Embodiment 1 is identical;

[0059] (4) Locating the target brain area: first glue the wound with biologic...

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PUM

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Abstract

The invention relates to a monkey virus injection method and applications thereof, and more specifically relates to a monkey encephalic zone virus injection method, and applications thereof. The monkey encephalic zone virus injection method mainly comprises following steps: virus is coated so as to realize coupling of exposed surface residues on virus housings with magnetic induction proteins, motor proteins, and ATP, and obtaining of modified virus (nanometer robots), and spinal puncture technology is adopted for injection into monkey spinal fluid; and at last, under external magnetic field control, guiding of the modified virus in a target encephalic zone is carried out. Compared with the prior art, the advantages are that: no damage on monkey brain is caused, activation of immunity reaction of monkey brains is avoided, virus release is realized, and at the same time, virus expression efficiency in monkey brains is increased. Under cooperation of magnetic induction proteins and computer controlled magnetic force, the modified virus can be dispersed, expression in encephalic zones of relatively large area is realized, and virus expression efficiency is increased greatly.

Description

technical field [0001] The invention relates to the field of non-human primate model animal virus research, in particular to a monkey virus injection method and application, in particular to a monkey brain region virus injection method and application. Background technique [0002] As a non-human primate model animal, monkeys are often used in the study of human diseases, but there is still no breakthrough in the research on neural circuit diseases. The main reason is that the tools and viruses used to study neural circuits are difficult brain expression. The traditional method of virus injection in the monkey brain area is similar to that of rodents. First, the monkey is anesthetized with a mixed anesthetic; second, after routine disinfection, the monkey’s scalp is cut open, the skull is preserved, the monkey brain is fixed, and the zygomatic arch and positive The mid-sagittal suture was defined as a bony landmark, and the injection site was determined with reference to th...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61D7/00C12Q1/70
CPCA61D7/00C12N7/00C12Q1/68C12Q1/70
Inventor 贾香连李宁宁刘晓雷路中华王立平
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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