Staff:
Pina Marotta
Maddalena Pizzulo
Maria Marotta
Filomena Russo |
Genetically modified mouse model are a useful tool to dissect biological processes both normal and/or pathological. Recent techniques pushed Genome Modification. Mouse genome can be easily engineered allowing DNA targeting: promoter, gene, miRNA etc.
In general, such improvement changed the way to study phenotype from forward to reverse genetic.
Biogem offers quality custom cell line and mouse model generation through:
- Gene targeting vector.
- Embryonic Stem (ES) cells modification.
- ES cells for homologous recombination.
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| KNOCK OUT: |
| CONSTITUTIVE |
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A Constitutive Knock Out will eliminate the function of the targeted gene in all cells throughout development and adulthood. Traditional knock-out involves deleting or disabling both copies of a specific gene. It is indicated for study of gene function, drug and therapeutic development and disease research.
The construction of this targeting vector include the following major steps: |
| CONDITIONAL |
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Conditional knock-out involves deleting or disabling a gene in only a particular organ, tissue, or cell type or only during a certain development stage.
These system utilizes the Cre-lox system. This is a site-specific DNA recombination through the interaction of the Cre recombinase enzyme and loxP sites in the DNA strand: a mouse bearing the recombinase-specific sites is bred with a mouse expressing the recombinase. The tissue-specific expression of the recombinase allows the inactivation of the gene of interest only in the tissue where the recombinase is expressed.
Conditional knock-outs provide a temporal and tissue-specific gene knock-out control while allowing the mouse model to mature under the normal operating role of the gene of interest.
The construction of this targeting vector include the following major steps: |
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| GENE OVEREXPRESSION: |
| RANDOM INTEGRATION |
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This approach is based on random integration of the transgene in the genome by 'pronuclear microinjection' of the foreign DNA directly into the mouse egg just after fertilisation.
The final characteristics, such as expression pattern and level of expression, and relevance of the transgenic mouse obtained are unpredictable, more over all the research work must be performed on several independent lines.
The construction of this vector include the following major steps:
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| TARGETED INSERTION |
This approach, called "knock-in", is based on targeted insertion of the transgene in a well-characterized, transcriptionally active locus in the mouse genome, such as ROSA26 locus.
The construction of this targeting vector include the following major steps: |
| HUMANIZATION |
Humanization, based on Knock-in approach, allows for in vivo testing of compounds and antibodies against human proteins in the mouse. This is achieved by the replacement of the murine gene with the human counterpart, placing the gene in the same transcriptional context as the murine gene.
The construction of this targeting vector include the following major steps: |
| POINT MUTATIONS |
This approach involves the introduction of one or more point mutations anywhere in the target gene. This method can also be combined with conditional approaches, providing a temporal and tissue-specific gene mutation.
The construction of this targeting vector include the following major steps: |
| REPORTER FUSIONS |
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This targeting vector allows the introduction of a reporter gene to follow gene expression.
The construction of this targeting vector include the following major steps:
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| CONSTITUTIVE/INDUCIBLE CRE-DRIVER MOUSE STRAIN |
| The conditional approaches based on the Cre/lox system need mouse strains expressing the recombinase in a tissue- and time-specific manner, to make this models it is necessary the generation of a mouse that express Cre recombinase (constitutive or inducible) under the control of a specific promoter. The construction of this targeting vector include the following major steps: |
