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?

Key Concepts

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 introduces targeted mutations. By studying organisms with these alleles, researchers can infer the gene's function based on the phenotypic changes observed.

Gene Editing Techniques

Gene editing techniques, such as CRISPR-Cas9, allow scientists to make precise alterations to an organism's DNA. This technology utilizes a guide RNA to direct the Cas9 enzyme to a specific location in the genome, 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 can disrupt gene function, thus creating loss-of-function alleles.

Functional Genomics

Functional genomics is the field of study that aims to understand the relationship between genes and their functions within an organism. It employs various techniques, including gene knockout and overexpression studies, to elucidate the roles of genes, especially those with unknown functions. By analyzing the effects of loss-of-function alleles, researchers can gain insights into the biological pathways and processes in which these genes are involved.