High Throughput Cloning of Mouse Full-length cDNAs

点击翻译按钮获取中文摘要

DESCRIPTION (provided by applicant): Full length cDNA clones are the most powerful tools for the functional analyses of genes. It remains difficult, however, to isolate a physical cDNA clone from complex cDNA libraries, especially for long or rare transcripts. Large-scale cDNA and genomic sequencing has resulted in thousands of reference sequences in the NCBI database. While the cDNA sequences and genome sequencing provide important structural information, accession to a full-length clone remains the key if 1 is to proceed to study gene function or protein interactions. Many of the compiled gene sequences in the public databases do not have physical full-length clones available to support further investigation. OriGene Technologies, Inc. has developed over 20 state-of-the-art cDNA libraries (including 6 mouse cDNA libraries), the Rapid-Screen Arrayed cDNA Library Panels, to support high-throughput full-length cDNA cloning. To date, we have released 24,000 unique human full-length cDNA clones, the largest in the industry, for functional genomics research. Due to its genetic tractability and biological and pathological similarities between human and mouse, the mouse has emerged as a major model organism for basic mammalian biology and human diseases. Many experimental manipulations in mouse require a set of mouse full-length cDNA clones. Phase I of this project aims to isolate and sequence 50,000 mouse cDNA clones enriched with large inserts (>4 kb) in our RapidScreen cDNA libraries. This will include the identification and confirmation of new full-length cDNA clones when compared with existing mouse clone collections such as MGC and RIKEN. Phase II of the project will focus on the large-scale sequencing and identification of new full-length cDNA clones via sequencing up to 500,000 clones. Our ultimate goal is to produce at least 4000-5000 distinct mouse full-length cDNA clones (especially cDNAs with an ORF>3 kb) not currently part of the MGC collection. Our technology platform has proven to be both reliable and innovative, and the outcome of this project will assuredly facilitate current and future mouse functional genomics research.