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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.
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...
Gene Therapy00:59

Gene Therapy

Gene therapy is a technique where a gene is inserted into a person’s cells to prevent or treat a serious disease. The added gene may be a healthy version of the gene that is mutated in the patient, or it could be a different gene that inactivates or compensates for the patient’s disease-causing gene. For example, in patients with severe combined immunodeficiency (SCID) due to a mutation in the gene for the enzyme adenosine deaminase, a functioning version of the gene can be inserted. The...
Gene Therapy00:59

Gene Therapy

Gene therapy is a technique where a gene is inserted into a person’s cells to prevent or treat a serious disease. The added gene may be a healthy version of the gene that is mutated in the patient, or it could be a different gene that inactivates or compensates for the patient’s disease-causing gene. For example, in patients with severe combined immunodeficiency (SCID) due to a mutation in the gene for the enzyme adenosine deaminase, a functioning version of the gene can be inserted. The...
Combinatorial Gene Control02:33

Combinatorial Gene Control

Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...

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Related Experiment Video

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

[Issues and solutions of conditional gene targeting].

Fu-Yu Lin1, Xiao Yang

  • 1Genetic Laboratory of Development and Diseases, State Key Laboratory of Proteomics, Institute of Biotechnology, Beijing 100071, China. linfuyu2000@yahoo.com.cn

Yi Chuan = Hereditas
|May 19, 2011
PubMed
Summary

Conditional gene targeting using Cre-loxP is crucial for studying gene function and creating disease models. Addressing Cre transgene expression, efficiency, and toxicity issues ensures accurate genetic research and mouse model interpretation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Context:

  • Conditional gene targeting, particularly using the Cre-loxP system, is fundamental for dissecting gene function and developing disease models in mice.
  • However, the reliability of this system can be compromised by factors like unintended Cre transgene expression, variable recombination efficiency, and Cre recombinase toxicity.

Purpose:

  • To highlight potential pitfalls in the Cre-loxP system that can affect the accuracy of gene function studies and phenotypic analysis in genetically engineered mice.
  • To emphasize the importance of addressing these challenges for robust and reproducible research outcomes.

Summary:

  • The Cre-loxP system, while powerful, faces challenges including aberrant Cre expression, inconsistent recombination rates, and potential toxicity.
  • These issues, along with genetic background, breeding strategies, and experimental controls, can significantly impact the interpretation of gene function and disease phenotypes.
  • Implementing strategies for controlled Cre expression, efficiency assessment, toxicity mitigation, and optimized experimental design is crucial.

Impact:

  • Ensuring the accuracy of gene functional studies and phenotypic interpretation in conditional gene targeting mouse models.
  • Improving the reliability and reproducibility of research utilizing the Cre-loxP system.
  • Facilitating the generation of more precise and informative disease models for biomedical research.