Classical and contemporary approaches for establishing gene function</h5> With the development of new and affordable methods for whole-genome sequencing, and the design and implementation of large genome annotation projects, scientists are poised to deliver upon the promises of the genomic revolution to transform basic science and personalized medicine. The resulting wealth of information presents researchers with a new primary challenge of converting this enormous amount of data into functionally and clinically relevant knowledge. Central to this problem is the need for efficient and reliable methods that enable investigators to determine how genotype influences phenotype. Targeted gene inactivation via homologous recombination is a powerful method capable of providing conclusive information for evaluating gene function  . However, the use of this technique has been hampered by several factors, including the low efficiency at which engineered constructs are correctly inserted into the chromosomal target site, the need for time-consuming and labor-insensitive selection/screening strategies, and the potential for adverse mutagenic effects. Targeted gene knockdown by RNAi (see Glossary ) has provided researchers with a rapid, inexpensive, and high-throughput alternative to homologous recombination  . However, knockdown by RNAi is incomplete, varies between experiments and laboratories, has unpredictable off-target effects, and provides
Trends in Biotechnology – Elsevier
Published: Jul 1, 2013
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