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Related Concept Videos

Combinatorial Gene Control02:33

Combinatorial Gene Control

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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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Genetic Screens02:46

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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
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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.
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Related Experiment Video

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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
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A versatile modular vector system for rapid combinatorial mammalian genetics.

Joachim Albers, Claudia Danzer, Markus Rechsteiner

    The Journal of Clinical Investigation
    |March 10, 2015
    PubMed
    Summary
    This summary is machine-generated.

    The Multiple Lentiviral Expression (MuLE) system enables simultaneous genetic modifications in mammalian cells. This powerful tool accelerates the engineering of complex tumors and disease models for research.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Cancer Research

    Background:

    • Introducing multiple genetic alterations into cells is crucial for understanding complex diseases like cancer.
    • Existing methods can be time-consuming and lack flexibility for combinatorial genetic engineering.

    Purpose of the Study:

    • To develop and present the Multiple Lentiviral Expression (MuLE) system for efficient, simultaneous genetic engineering.
    • To demonstrate the MuLE system's utility in creating complex polycistronic lentiviruses for various applications.

    Main Methods:

    • Development of a modular toolbox of MuLE vectors.
    • Utilizing MuLE vectors for gene overexpression, knockdown, deletion, and CRISPR/Cas9-mediated mutation.
    • Incorporating fluorescent or enzymatic reporters for cellular assays and animal imaging.
    • Application of the MuLE system in tumor engineering models.

    Main Results:

    • Engineered tumors with up to 5 genetic alterations in primary mouse cells.
    • Successfully identified genetic dependencies and performed genetic interaction screens.
    • Generated 3 murine sarcoma models by simultaneously knocking out tumor suppressor genes.
    • Demonstrated rapid generation and monitoring of genetically defined autochthonous tumors.

    Conclusions:

    • The MuLE system offers a flexible and modular approach for rapid combinatorial genetic engineering.
    • It significantly accelerates the creation of complex cellular and animal models for disease research.
    • The system provides powerful genetic tools for systematically investigating disease mechanisms.