You have cloned the mouse ortholog (see Genetic Analysis 14.2 for definition) of the gene associated with human Huntington disease (HD) and wish to examine its expression in mice. Outline the approaches you might take to examine the temporal and spatial expression pattern at the cellular level.

Verified step by step guidance

1. Step 1: Use in situ hybridization to detect mRNA expression. This technique allows you to visualize where and when the gene is being expressed in tissue sections by using a labeled complementary RNA or DNA probe.
2. Step 2: Perform immunohistochemistry to detect protein expression. This involves using antibodies specific to the protein encoded by the gene to determine its presence and distribution in tissue samples.
3. Step 3: Utilize reporter gene assays. Introduce a reporter gene, such as GFP (Green Fluorescent Protein), under the control of the gene's promoter to visualize expression patterns in live tissues or organisms.
4. Step 4: Conduct quantitative PCR (qPCR) to measure mRNA levels. This technique provides quantitative data on gene expression levels at different developmental stages or in different tissues.
5. Step 5: Analyze expression using RNA sequencing (RNA-seq). This high-throughput method allows for a comprehensive analysis of gene expression across different tissues and developmental stages, providing insights into temporal and spatial patterns.

Key Concepts

Gene Cloning

Gene cloning is a molecular biology technique used to create copies of a specific gene. In this context, the mouse ortholog of the gene associated with Huntington disease has been cloned, allowing researchers to study its function and expression. This process typically involves isolating the gene of interest, inserting it into a vector, and introducing it into host cells for propagation.

Gene Expression Analysis

Gene expression analysis involves measuring the activity of a gene to understand its role in cellular processes. Techniques such as quantitative PCR, in situ hybridization, and RNA sequencing can be employed to assess both the temporal (when) and spatial (where) expression patterns of the cloned gene in mouse tissues. This information is crucial for linking gene function to specific biological processes and disease mechanisms.

Temporal and Spatial Expression Patterns

Temporal and spatial expression patterns refer to the timing and location of gene expression within an organism. Temporal expression examines how gene activity changes over time, while spatial expression focuses on where in the body the gene is active. Understanding these patterns is essential for elucidating the role of the Huntington disease gene in development and pathology, particularly in relation to the onset and progression of the disease.