Molecular Genetics: Gene Duplication and Mutation
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Questions and Answers

What is the significance of using the C2H2 zinc finger motif as a platform for designing DNA binding modules with novel specificities?

  • It eliminates the risk of insertional mutagenesis and artefacts driven by variegated expression levels
  • It allows for the replacement of an endogenous gene with an overexpressed copy
  • It enables gene editing without the need for gene targeting
  • It has enabled the production of novel ZFPs against investigator-specified loci (correct)
  • What is the benefit of using ZFNs in gene targeting?

  • It eliminates the risk of artefacts driven by variegated expression levels
  • It eliminates the need for allele engineering
  • It allows for bona fide reverse genetics in diverse model organisms (correct)
  • It forces the use of surrogate genes
  • What is the advantage of using ZFNs over traditional gene targeting methods?

  • It requires variegated expression levels
  • It allows for the use of cDNA transgenes
  • It enables the use of surrogate genes
  • It eliminates the risk of insertional mutagenesis events (correct)
  • What is the significance of allele engineering in somatic cell genetics?

    <p>It is enabled by ZFNs</p> Signup and view all the answers

    What is the benefit of using current-generation ZFNs in gene-function studies?

    <p>It removes constraints on experimental design</p> Signup and view all the answers

    What is the significance of tagging an endogenous gene with GFP?

    <p>It enables the simultaneous elimination of artefacts driven by variegated expression levels</p> Signup and view all the answers

    What is the significance of using ZFNs in cell-based therapy?

    <p>It enables the production of novel ZFNs against investigator-specified loci</p> Signup and view all the answers

    What is the significance of ZFNs in gene targeting?

    <p>They enable the production of novel ZFPs against investigator-specified loci</p> Signup and view all the answers

    What is the primary outcome of using ZFNs to introduce small insertions and/or deletions at the site of the break?

    <p>Disruption of a target gene</p> Signup and view all the answers

    What is the role of an investigator-designed homologous donor DNA in gene correction?

    <p>To provide a template for repair</p> Signup and view all the answers

    What is the result of ZFPs interacting with neighbouring fingers and bases outside their proximal DNA triplet?

    <p>Complicated efforts to produce new ZFPs</p> Signup and view all the answers

    What is the outcome of using ZFNs in an embryo, resulting in up to 10% of the progeny being mutated for the gene of interest?

    <p>Gene disruption</p> Signup and view all the answers

    What is the purpose of modular design in making ZFPs with new sequence specificities?

    <p>To enable more selective binding</p> Signup and view all the answers

    What is the result of duplication events, usually with a subsequent divergence of function?

    <p>Formation of functional paralogues with new sequence specificities</p> Signup and view all the answers

    What is the role of ZFNs in gene correction, in the presence of an investigator-designed homologous donor DNA?

    <p>To repair a double-strand break</p> Signup and view all the answers

    What is the characteristic of non-functional paralogues, also known as pseudogenes?

    <p>Lack of promoter or intron sequences</p> Signup and view all the answers

    What is the primary goal of creating a point mutation in a gene?

    <p>To study gene function and/or model disease-causing mutations</p> Signup and view all the answers

    What is the advantage of using isogenic settings in gene-function studies?

    <p>It provides a way to study structure–function relationships at native levels of expression</p> Signup and view all the answers

    What is the term for the process of creating a new allele of a gene?

    <p>Allele engineering</p> Signup and view all the answers

    What is the main challenge in genetically engineering human stem cells?

    <p>Their recalcitrance to genetic engineering</p> Signup and view all the answers

    What is the term for the study of the function of a gene by creating a mutation in the gene?

    <p>Reverse genetics</p> Signup and view all the answers

    What is the application of gene targeting in human ES and iPS cells?

    <p>Modelling of human tissues (diseased and normal) in basic science, drug discovery, and regenerative medicine</p> Signup and view all the answers

    What is the advantage of using ZFNs in gene targeting?

    <p>They enable the creation of point mutations at a specific position in the target gene</p> Signup and view all the answers

    What is the term for the creation of a new allele of a gene through the correction of a point mutation?

    <p>De novo creation of point mutations</p> Signup and view all the answers

    Study Notes

    Zinc Finger Proteins (ZFPs) and Zinc Finger Nucleases (ZFNs)

    • ZFPs can be designed to bind specific DNA sequences through modular design, but this can be complicated by interactions between fingers and neighboring base pairs.
    • ZFNs can introduce small insertions and/or deletions at the site of a break, allowing for gene disruption or correction.
    • ZFNs can be used to create point mutations in a target gene, mimicking disease-causing mutations or disabling motifs crucial for function.

    Methods for Producing Novel ZFPs

    • Duplication events can create new ZFPs with novel specificities through subtle divergence of function.
    • Alternative strategies for making ZFPs include modular design, which can be complicated by interactions between fingers.

    Gene Correction and Addition

    • ZFNs can be used to correct point mutations in a target gene, allowing for the study of gene function and disease-causing mutations.
    • ZFNs can be used to create isogenic panels of mouse ES cells carrying defined alleles for an endogenous gene.
    • Gene addition in human ES and iPS cells can be used to model human tissues and study structure-function relationships.

    Applications of ZFNs

    • ZFNs have been used to create point mutations in D.melanogaster, with up to 90% of treated animals yielding offspring with donor-specified alleles.
    • ZFNs can be used to study gene function and disease-causing mutations in human stem cells.
    • The application of ZFNs removes many constraints on experimental design, allowing for more precise and efficient gene editing.

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    Description

    This quiz covers the concepts of gene duplication, mutation, and genetic variation, including the process of gene duplication events and the potential outcomes.

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