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Adeno-associated virus with variant capsid protein and application thereof

A capsid protein and virus technology, applied in the field of adeno-associated virus, can solve problems such as unsuitable for continuous treatment

Pending Publication Date: 2020-10-27
INNOVATION ACAD FOR PRECISION MEASUREMENT SCI & TECH CAS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, AAVs with both these important features have not been reported
Retrograde infection of AAV is mainly achieved through intracranial injection to achieve most of the cell infection, which is not as convenient as intravenous injection, but antibodies may be produced after intravenous injection, which is not suitable for continuous treatment. Therefore, the development of multi-channel targeting brain neurons adeno-associated virus

Method used

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  • Adeno-associated virus with variant capsid protein and application thereof
  • Adeno-associated virus with variant capsid protein and application thereof

Examples

Experimental program
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Embodiment Construction

[0016] 1. Construction of AAV variant (named AAV9-RB) Rep-Cap plasmid

[0017] The serotype of AAV is determined by the sequence of the Cap protein in the Rep-Cap plasmid required for AAV packaging, and different serotypes of AAV have different tissue tropisms. Therefore, new AAV viruses can be obtained by modifying the Cap protein. The DNA sequence (SEQ ID NO.2) encoding LADQDYTKTA (SEQ ID NO.1) was inserted between amino acids 588 and 589 of the VP1 capsid protein of the parent AAV9, and the specific process included the following aspects:

[0018] ① Using the parental AAV9 plasmid pAAV-RC9 (purchased from Wuhan Privy Brain Science and Technology Co., Ltd.) as a template, use Takara Primerstar Polymerase (Takara Company) to amplify the Retro1 fragment. The sequence of the primer Retro1-F is shown in SEQ ID NO.3, The sequence of primer Retro1-R is shown in SEQ ID NO.4; the PCR reaction system is 50 μl: 5×Reaction Buffer 10 μl, 10 mM dNTPs 1 μl, 10 μM forward primer 2.5 μl, 1...

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Abstract

The invention discloses an adeno-associated virus with a variant type capsid protein and application thereof. The adeno-associated virus with the variant type capsid protein AAV9-RB is obtained by inserting an amino acid sequence LADQDYTKTA between Q588 and A589 of capsid protein VP1 of wild type AAV9. The adeno-associated virus with the variant type capsid protein AAV9-RB is the first reported virus body that can not only effectively reverse infection from the axon terminal, but also cross the blood-brain barrier to target brain cells. The invention provides better technical support for neuroscience studies and brain disease treatments, with wide application value and prospect.

Description

technical field [0001] The invention belongs to the technical field of adeno-associated virus, and in particular relates to an adeno-associated virus with mutated capsid protein and application thereof. Background technique [0002] Viral vectors, especially those that allow efficient gene transfer from axon terminals or across the blood-brain barrier to the central nervous system (CNS), are useful for analyzing the structure and function of specific neural circuits around injection sites and can be Gene delivery to distant target regions has become one of the most potential and promising therapeutic tools. Compared with traditional retrograde tracers, viral vectors can express genes in specific neuronal populations, such as optogenetic, chemogenetic and calcium-sensitive functional probes, which are widely used to monitor and manipulate neuronal activities. Several natural and engineered neurotropic viruses exhibit retrograde infection capabilities, including pseudorabies ...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N7/01C12N15/864A61K9/00A61K47/42A61K47/46A61K48/00A61P25/00C12R1/93
CPCC07K14/005C12N7/00C12N15/86A61K48/00A61K9/0019A61K9/0085A61K47/46A61K47/42A61P25/00C07K2319/00C12N2750/14122C12N2750/14143Y02A50/30
Inventor 林坤章徐富强
Owner INNOVATION ACAD FOR PRECISION MEASUREMENT SCI & TECH CAS
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