Transgenic Mice -- overexpression of the target gene

The mouse strains obtained by experimentally introducing exogenous genes into early mouse embryos and integrating them into the genome, which can stably inherit the exogenous genes.

Random Insertion

Random insertion transgenesis
Transposon-mediated site-selective transgenesis

Site-specific Insertion

ES/Cas9-mediated site-specific knock-in transgenesis
Conditional overexpression

Random Transgensis

Plasmid construction

Microinjection

Embryo transfer

Obtain F0

Advantages of random transgenesis

Simple construction, low technical difficulty
Short cycle（obtain F0 in 3 months）
Allows for the insertion of large fragments, enabling transgenic models with over 100kb

Disadvantages of random transgenesis

random insertion leads to uncertain expression or expression intensity; uncertain expression type.
requires establishing lines, which is time-consuming.
at least 2-3 expressing lines are needed to validate phenotypes.

Site-specific transgenesis

Insert exogenous DNA at a 'Safe Harbor' site that allows for safe knock-in and ensures stable expression of the target gene, thereby obtaining site-specific transgenic mice.

Random transgenesis

Low technical difficulty, short cycle
Insertion site, copy number random
Likely to disrupt other endogenous genes
Substantial effort required for later line establishment and screening

Targeted transgenesis

Obtaining a unique model, does not affect the expression of endogenous genes
High genetic stability
Analysis possible without the need for screening

Direct overexpression

Direct overexpression Knock-in target gene fragment at Safe Harbor sites Safe Harbors：Rosa26、H11、Col1a1 etc.

-Rosa26： located on mouse chromosome 6
-H11： located on mouse chromosome 11
-Col1a1： located on mouse chromosome 6

Conditional overexpression

In the construction of site-specific conditional overexpression mouse models, the Cre-LoxP system is used to regulate inducible expression of the target gene.

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