TABLE 7-2 Transgenic Technologies Commonly Used in Laboratory Mice

Method

Purpose

DNA Construct

Desired Effect

Variations

DNA microinjection into zygotic pronucleus

Overexpression

Gene of interest with strong promoter

Higher than normal expression

Tissue specific, ubiquitous or inducible promoter

Misexpression

Gene of interest with ectopic promoter

Ectopic expression

Temporal or spatial misexpression

Promoter and enhancer analysis

Regulatory regions driving reporter gene expression

Mapping of regulatory elements

Various reporter genes (e.g., lacZ), green fluorescent protein, luciferase

Antisense

Endogenous promoter with antisense gene

Reduced expression of endogenous gene

 

Insertional mutagenesis

Any

Phenotype associated with serendipitous disruption of unknown gene

 

Gene trapping

Promoterless reporter gene

Expression of reporter gene indicating expression of endogenous gene

Various reporter genes (e.g., lacZ), green fluorescent protein, luciferase

Targeted mutagenesis via homologous recombination in embryonic stem cells

 

DNA electroporation into embryonic stem cells

Mutations

Homologous genomic regions flanking selectable marker

Mutation of endogenous gene

Insertion into or deletion of endogenous genomic sequences to produce null (knockout), hypomorph, or dominant negative mutations



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