Intro practical prep
20 Questions
2 Views

Choose a study mode

Play Quiz
Study Flashcards
Spaced Repetition
Chat to lesson

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

What is the purpose of adding additional bases at the 5' ends of both oligos?

  • To enhance the efficiency of the Cas9 protein
  • To increase the on-target score of the gRNA
  • To reduce the likelihood of off-target effects
  • To ensure the DNA oligos can be cloned into a vector (correct)

    • What is the advantage of designing a complementary strand and flipping it to make the reverse strand?

  • It reduces the specificity of the gRNA
  • It increases the likelihood of Cas9 DNA cleavage (correct)
  • It increases the risk of off-target effects
  • It reduces the binding efficiency of the gRNA to the target site

    • What is the format of the cDNA sequence used to design the gRNA?

  • GenBank format
  • SBOL format
  • GFF format
  • FASTA format (correct)

    • What is the purpose of using IDT to get custom gRNA?

    <p>To get a selection of target sequences with on-target scores</p> Signup and view all the answers

    What is the purpose of using a bacterial expression plasmid containing a cDNA for fluorescent protein?

    <p>To allow for fluorescence when transformed into bacteria</p> Signup and view all the answers

    What is the primary function of the Cas9 protein in the described experiment?

    <p>To cut out specific DNA sequences</p> Signup and view all the answers

    What is the purpose of using a complementary strand and flipping it to make the reverse strand?

    <p>To ensure efficient targeting of both strands of DNA</p> Signup and view all the answers

    What type of sequence is used to design the gRNA?

    <p>cDNA sequence</p> Signup and view all the answers

    What is the purpose of adding the 'atag' sequence to the forward oligo?

    <p>To facilitate cloning into a vector</p> Signup and view all the answers

    What is the purpose of using IDT to get custom gRNA?

    <p>To obtain a selection of target sequences with on-target scores</p> Signup and view all the answers

    What is the expected outcome when a bacterial expression plasmid containing a cDNA for fluorescent protein is transformed into bacteria?

    <p>The bacteria will fluoresce</p> Signup and view all the answers

    What is the significance of the on-target score provided by IDT for the target sequences?

    <p>It predicts the efficacy of the gRNA in gene knockout</p> Signup and view all the answers

    What is the purpose of designing a DNA oligo for the Cas9 protein?

    <p>To create a custom gRNA for gene knockout</p> Signup and view all the answers

    What is the advantage of using a cDNA sequence for fluorescent protein in the design of the gRNA?

    <p>It provides a template for the design of the gRNA</p> Signup and view all the answers

    What is the expected outcome of using the Cas9 protein to cut out the fluorescent protein gene?

    <p>The fluorescent protein gene will be knocked out</p> Signup and view all the answers

    What is the primary purpose of using a fluorescent protein in the experiment?

    <p>To identify potential targets for the gRNA</p> Signup and view all the answers

    What is the significance of using both forward and reverse strands in the design of the DNA oligo?

    <p>It ensures efficient targeting of the gene by the Cas9 protein</p> Signup and view all the answers

    What is the role of the on-target score provided by IDT?

    <p>It selects the optimal target sequence for the gRNA</p> Signup and view all the answers

    What is the purpose of transforming the bacterial expression plasmid into bacteria?

    <p>To express the fluorescent protein in the bacteria</p> Signup and view all the answers

    What is the ultimate goal of designing gRNAs to knockout fluorescent protein genes?

    <p>To create a bacterial strain that does not express the fluorescent protein</p> Signup and view all the answers

    Study Notes

    Bacterial Expression Plasmids

    • Contain cDNAs for fluorescent proteins (EGFP or mCherry), leading to fluorescence when transformed into bacteria

    gRNA Design

    • Used IDT to design custom gRNA for chosen protein
    • Imported cDNA sequence in FASTA format
    • Received a selection of target sequences from IDT, each with an on-target score

    Oligo Design for Cas9 Protein

    • Used target sequence as forward strand
    • Designed complementary strand and flipped it to make the reverse strand for efficient targeting
    • Added additional bases at the 5’ ends of both oligos for cloning into a vector
      • Forward oligo: added atag
      • Reverse oligo: added aaac

    Practical Overview

    • Designed gRNAs to knock out fluorescent protein genes using Cas9 protein

    Studying That Suits You

    Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

    Quiz Team

    Description

    This quiz covers the design of bacterial expression plasmids, gRNA design, and oligo design for Cas9 protein. Learn about CRISPR-Cas9 gene editing and its applications in molecular biology.

    More Like This

    CRISPR/Cas9 Gene Editing
    5 questions

    CRISPR/Cas9 Gene Editing

    SofterFuchsia avatar
    SofterFuchsia
    CRISPR-Cas9 and Tissue-Specific Genome Editing Quiz
    23 questions

    CRISPR-Cas9 and Tissue-Specific Genome Editing Quiz

    GallantSnowflakeObsidian avatar
    GallantSnowflakeObsidian
    CRISPR/CAS9 Gene Editing Tool
    24 questions

    CRISPR/CAS9 Gene Editing Tool

    FineLookingPink avatar
    FineLookingPink
    CRISPR-Cas9: Sistema de edición genética
    20 questions

    CRISPR-Cas9: Sistema de edición genética

    MagicalClematis avatar
    MagicalClematis
    Use Quizgecko on...
    Browser
    Open
    Browser
    Browser