Gene Function Analysis

Introduction

This volume of Methods in Molecular Biology focuses on techniques to determine the function of a gene. Traditionally, the function of a gene was determined following cloning, which provided its DNA sequence and an ability to modify this sequence.

Experiments were performed that looked for phenotypic changes in a cell line or model organism following modifications to the sequence, knocking out of the gene, or enhancing expression of the gene.

Advancements in Gene Identification

In the 1990’s, the growing sequence databases and the BLAST algorithm provided additional power by allowing identification of genes with known function that had similar sequences and potentially similar molecular mechanisms.

Experimental Methods

On the experimental side, methods, such as two-hybrid screening that could directly determine the partners of specific proteins and even the domains of interaction, came into widespread use.

High-Throughput Technologies and Challenges

With the advent of high-throughput technologies following completion of the human genome project and similar projects in model organisms, the number of genes of interest has expanded and the traditional methods for gene function analysis cannot achieve the throughput necessary for large-scale exploration.

Although computational tools such as BLAST remain a good point of departure, it is often the case that a gene that appears interesting in a high-throughput experiment shows no obvious similarity to a gene of known function.

In addition, when BLAST does find a similar gene, the process has often only begun.