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Introducing a specific change in the mouse genome using CRISPR/Cas9 requires: a. Alterations to both alleles of a gene b. An HDR gene fragment c. Non-homologous end joining (NHEJ) d. Leaving out the guide RNA e. All of the choices

Introducing a specific change in the mouse genome using CRISPR/Cas9 requires: a. Alterations to both alleles of a gene b. An HDR gene fragment c. Non-homologous end joining (NHEJ) d. Leaving out the guide RNA e. All of the choices

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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### Introducing a Specific Change in the Mouse Genome Using CRISPR/Cas9 CRISPR/Cas9 technology allows for precise modifications in the genome. When introducing a specific change in the mouse genome using CRISPR/Cas9, it requires: - **a. Alterations to both alleles of a gene:** This involves making changes to both copies of the gene present in the organism to ensure the modification is thorough and can be passed on to future generations. - **b. An HDR gene fragment:** HDR (Homology-Directed Repair) is a pathway for DNA repair. Including an HDR gene fragment can help guide the repair mechanism to incorporate the intended changes. - **c. Non-homologous end joining (NHEJ):** NHEJ is another DNA repair pathway that can join DNA ends after a break, often leading to insertions or deletions (indels) at the repair site. - **d. Leaving out the guide RNA:** This would mean not including the RNA sequence that directs the Cas9 enzyme to the specific location in the genome. Note that leaving out the guide RNA would typically render the CRISPR/Cas9 system ineffective. - **e. All of the choices:** This option suggests that all of the above conditions are required, which is not the case for the proper functionality of CRISPR/Cas9. ### Explanation of Components: 1. **Alterations to Both Alleles:** Ensuring that both alleles are altered guarantees that the genetic modification is homozygous. This is especially important in studies where the recessive trait needs to be expressed or when ensuring the trait is inherited by offspring. 2. **HDR Gene Fragment:** This is used when precise modifications are needed, such as inserting or correcting a gene sequence. The HDR pathway leverages a template with the desired change to accurately repair the break induced by the Cas9 enzyme. 3. **NHEJ:** This is a more error-prone repair mechanism that the cell naturally uses to quickly fix double-strand breaks. It is useful for knocking out genes by causing frameshift mutations. 4. **Guide RNA (gRNA):** This small piece of RNA is crucial because it directs the Cas9 enzyme to the correct location in the genome. Without it, Cas9 would not know where to create the DNA break. 5. **Choosing the Correct Option:** The
Transcribed Image Text:### Introducing a Specific Change in the Mouse Genome Using CRISPR/Cas9 CRISPR/Cas9 technology allows for precise modifications in the genome. When introducing a specific change in the mouse genome using CRISPR/Cas9, it requires: - **a. Alterations to both alleles of a gene:** This involves making changes to both copies of the gene present in the organism to ensure the modification is thorough and can be passed on to future generations. - **b. An HDR gene fragment:** HDR (Homology-Directed Repair) is a pathway for DNA repair. Including an HDR gene fragment can help guide the repair mechanism to incorporate the intended changes. - **c. Non-homologous end joining (NHEJ):** NHEJ is another DNA repair pathway that can join DNA ends after a break, often leading to insertions or deletions (indels) at the repair site. - **d. Leaving out the guide RNA:** This would mean not including the RNA sequence that directs the Cas9 enzyme to the specific location in the genome. Note that leaving out the guide RNA would typically render the CRISPR/Cas9 system ineffective. - **e. All of the choices:** This option suggests that all of the above conditions are required, which is not the case for the proper functionality of CRISPR/Cas9. ### Explanation of Components: 1. **Alterations to Both Alleles:** Ensuring that both alleles are altered guarantees that the genetic modification is homozygous. This is especially important in studies where the recessive trait needs to be expressed or when ensuring the trait is inherited by offspring. 2. **HDR Gene Fragment:** This is used when precise modifications are needed, such as inserting or correcting a gene sequence. The HDR pathway leverages a template with the desired change to accurately repair the break induced by the Cas9 enzyme. 3. **NHEJ:** This is a more error-prone repair mechanism that the cell naturally uses to quickly fix double-strand breaks. It is useful for knocking out genes by causing frameshift mutations. 4. **Guide RNA (gRNA):** This small piece of RNA is crucial because it directs the Cas9 enzyme to the correct location in the genome. Without it, Cas9 would not know where to create the DNA break. 5. **Choosing the Correct Option:** The
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