VelociGene: a high-throughput approach for functionizing the genome via custom gene mutation and high-resolution expression analysis in mice

Now that the genome has been sequenced, determining gene function presents the next major challenge. Many scientists agree that the most powerful technologies for determining gene function involve genetic manipulations that knock out, replace, or overexpress gene products in mice so as to evaluate functional consequences. Unfortunately, even in the most sophisticated laboratories, such approaches still remain rather custom and low-throughput. I will describe a new set of technologies that allow for an unprecedented rate of generation of knockouts, knockins and transgenics–easily industrializable and scaleable to thousands per year. These approaches involve rapid manipulation of very large pieces of DNA (hundreds of kilobases in size), allowing the entire genome to be spanned by about 25,000 separate pieces of DNA. VelociGene has enormous flexibility, allowing the production of custom mutations with nucleotide precision, deletions of very large size, reporter knockins, transgenic overexpression, as well as conditional and complex alleles. Genetically modified mice produced via VelociGene are phenotyped using a variety of high-throughput approaches, ranging from high-throughput/high-resolution reporter gene analyses to localize the target gene with cellular resolution, to four-dimensional transcriptional fingerprinting via microarray. These approaches have identified and validated multiple targets in therapeutic areas such as obesity and diabetes, arthritis and cartilage growth, muscle atrophy, and angiogenesis and cancer.