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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
Retrovirus Life Cycles01:10

Retrovirus Life Cycles

Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the retrovirus to...
Viral Mutations00:36

Viral Mutations

A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material for adaptive...
Retroviruses02:33

Retroviruses

Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
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Retroviral insertional mutagenesis: past, present and future.

A G Uren1, J Kool, A Berns

  • 1Division of Molecular Genetics, Netherlands Cancer Institute, Amsterdam.

Oncogene
|November 22, 2005
PubMed
Summary

Retroviral insertion mutagenesis screens in mice efficiently identify oncogenic mutations. These findings are crucial for understanding human cancer development and developing new therapeutic strategies.

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Area of Science:

  • Oncology
  • Genetics
  • Molecular Biology

Background:

  • Retroviral insertion mutagenesis in mice is a key method for discovering oncogenes.
  • Many identified oncogenes are implicated in human cancers.
  • Advances in genome sequencing and insertion site cloning enhance oncogenic event identification.

Purpose of the Study:

  • To review the features of retroviral insertion mutagenesis screens.
  • To discuss mechanisms of retroviral mutagenesis, cloning techniques, and data analysis.
  • To explore the application of insertional mutagens in non-hematopoietic and non-mammary tumors.

Main Methods:

  • Review of existing literature on retroviral insertion mutagenesis.
  • Discussion of molecular mechanisms of insertional mutagenesis.
  • Analysis of techniques for insertion site cloning and common insertion site identification.

Main Results:

  • Retroviral insertions activate oncogenes through various mechanisms.
  • Improved cloning methods facilitate precise identification of oncogenic insertions.
  • Common insertion site analysis reveals frequently mutated oncogenes.

Conclusions:

  • Retroviral insertion mutagenesis is a valuable tool for in vivo oncogene discovery.
  • This approach aids in understanding human cancer etiology.
  • Future applications include studying diverse tumor types beyond hematopoietic and mammary cancers.