Transgénèse animale 2

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    Created by @ethel

    What are the steps in obtaining ES cells?

    Involves culturing, transferring Tg, injecting into blastocysts, and implantation.

    What is the consequence of using ES cells in transgenesis?

    Allows for in vitro selection of transgenic cell lines.

    What is a disadvantage of using ES cells for transgenesis?

    Requires a selection gene in the transgene that must be eliminated later.

    How does transgenesis by pronuclear transfer differ from using ES cells?

    Pronuclear transfer directly modifies zygotes, while ES cells involve multiple steps.

    What happens during gene transfer in gametes?

    Sperm are incubated with DNA to create transgenic animals.

    What is a challenge in stabilizing transgenic lines from gamete transfer?

    The transgene may become non-functional or rearranged.

    What is the significance of the 2007 Nobel Prize in Medicine?

    Recognized advancements in targeted gene manipulation techniques.

    What is the mechanism behind homologous recombination?

    Involves exchange between homologous DNA sequences.

    What is required for effective homologous recombination?

    Long homologous sequences and a rare event occurrence.

    What is the role of the targeting vector in gene targeting?

    Contains the gene of interest and homologous sequences for recombination.

    What is the role of the targeting vector in homologous recombination?

    It serves as a replacement or insertion vector for gene modification.

    How does the efficiency of homologous recombination vary with fragment size?

    Larger homologous fragments increase the frequency of recombination.

    What distinguishes type Ω homologous recombination from type O?

    Type Ω involves sequence replacement, while type O involves sequence insertion.

    What is the purpose of double selection in gene targeting?

    To enrich for recombinant ES cells by eliminating non-targeted integrations.

    What happens during the Cre/LoxP recombination process?

    It can lead to deletion, insertion, or inversion of DNA segments.

    What is a positive selection gene?

    It provides a survival advantage to cells by conferring drug resistance.

    What is a negative selection gene?

    It makes cells sensitive to a drug, thus eliminating them from the culture.

    What is the significance of using isogenic DNA in homologous recombination?

    It enhances the efficiency of gene targeting by reducing genetic variability.

    What is the consequence of targeted gene knockout?

    It results in the loss of protein expression from the targeted gene.

    What is the function of the recombinase in the Cre/LoxP system?

    It recognizes LoxP sites to facilitate DNA recombination events.