Table of contents

1. Introduction to Genetics51m
2. Mendel's Laws of Inheritance3h 37m
3. Extensions to Mendelian Inheritance2h 41m
4. Genetic Mapping and Linkage2h 28m
5. Genetics of Bacteria and Viruses1h 21m
6. Chromosomal Variation1h 48m
7. DNA and Chromosome Structure56m
8. DNA Replication1h 10m
9. Mitosis and Meiosis1h 34m
10. Transcription1h 0m
11. Translation58m
12. Gene Regulation in Prokaryotes1h 19m
13. Gene Regulation in Eukaryotes44m
14. Genetic Control of Development44m
15. Genomes and Genomics1h 50m
16. Transposable Elements47m
17. Mutation, Repair, and Recombination1h 6m
18. Molecular Genetic Tools19m
- Genetic Cloning
  9m
- Methods for Analyzing DNA
  9m
19. Cancer Genetics29m
- Overview of Cancer
  21m
- Cancer Mutations
  7m
20. Quantitative Genetics1h 26m
21. Population Genetics50m
- Hardy Weinberg
  19m
- Allelic Frequency Changes
  31m
22. Evolutionary Genetics29m

15. Genomes and Genomics

Functional Genomics

Genetics

15. Genomes and Genomics

Functional Genomics

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1:50 minutes

Problem 14a

Textbook Question

When the S. cerevisiae genome was sequenced and surveyed for possible genes, only about 40% of those genes had been previously identified in forward genetic screens. This left about 60% of predicted genes with no known function, leading some to dub the genes fun (function unknown) genes. As an approach to understanding the function of a certain fun gene, you wish to create a loss-of-function allele. How will you accomplish this?

Verified step by step guidance

Identify the target gene sequence: Obtain the DNA sequence of the fun gene from the S. cerevisiae genome database. This will allow you to design tools for gene disruption.

Design a disruption construct: Create a DNA construct that contains a selectable marker (e.g., an antibiotic resistance gene or a nutritional marker) flanked by sequences homologous to the regions upstream and downstream of the fun gene. These homologous regions will guide the construct to the correct location in the genome via homologous recombination.

Transform yeast cells: Introduce the disruption construct into S. cerevisiae cells using a transformation method such as electroporation or the lithium acetate method. This will allow the construct to enter the yeast cells.

Select for successful transformants: Grow the transformed yeast cells on a medium that selects for the presence of the selectable marker. Only cells where the disruption construct has integrated into the genome will survive.

Confirm the loss-of-function allele: Use molecular techniques such as PCR or Southern blotting to verify that the fun gene has been replaced by the disruption construct. Additionally, you can perform functional assays to confirm the loss of the gene's activity.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Loss-of-Function Alleles

Loss-of-function alleles are mutations that result in the complete or partial inactivation of a gene. These alleles can be generated through various methods, such as gene editing techniques like CRISPR-Cas9, which allows for precise modifications to the DNA sequence. By creating a loss-of-function allele, researchers can study the resulting phenotypic changes to infer the gene's role in biological processes.

Gene Editing Techniques

Gene editing techniques, such as CRISPR-Cas9, enable scientists to make targeted changes to an organism's genome. This technology utilizes a guide RNA to direct the Cas9 enzyme to a specific DNA sequence, where it creates a double-strand break. The cell's repair mechanisms then attempt to fix this break, often leading to insertions or deletions that disrupt gene function, thus facilitating the creation of loss-of-function alleles.

Functional Genomics

Functional genomics is the field of study that aims to understand the function of genes and their interactions within the genome. It employs various techniques, including gene knockout and overexpression studies, to elucidate the roles of specific genes in cellular processes. By investigating fun genes through functional genomics, researchers can uncover their biological significance and potential implications in health and disease.

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Related Practice

Textbook Question

Through a forward genetics screen in Arabidopsis you have identified a mutation that results in leaves curling upward, rather than being flat as in wild type. You have cloned the corresponding gene and note that it is a member of a small gene family composed of three additional members in Arabidopsis. How will you determine if the other three members of the gene family have similar or distinct functions as compared with the gene you first identified?

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Textbook Question

Transcription factors play key roles in the regulation of gene expression, but to do so, they must act within the nucleus. Like most proteins, however, transcription factors are translated in the cytoplasm. To enter the nucleus, transcription factors contain nuclear localization signals, which in some cases can work only when bound to some other molecule such as a steroid hormone. After entering the nucleus, transcription factors must bind to appropriate DNA sites and must interact with other transcription proteins at promoters, enhancers, and silencers. Transcription factors then activate or repress transcription through their activation or repression domains. Many drug therapies target transcription factors. Based on the information provided above, suggest three specific mechanisms through which a successful drug therapy, targeted to a transcription factor, might work.

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Textbook Question

The eyes of Drosophila develop from imaginal discs, groups of cells set aside in the fly embryo that differentiate into the adult structures during the pupal stage. Despite their importance in nature, eyes are dispensable for fruit fly life in the laboratory.

What complications might arise from genetic screens targeting an organ that differentiates late in development?

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Textbook Question

Devise a genetic screen to identify genes directing the development of the fly eye.

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Multiple Choice

Which of the following terms describes the study of physical interactions between DNA, RNA, and Protein?

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