High Throughput Cloning of Mouse Full-length cDNAs

描述（由申请人提供）：全长cDNA克隆是基因功能分析的最有力工具。然而，从复杂的cDNA文库中分离物理cDNA克隆仍然很困难，特别是对于长或罕见的转录本。大规模的cDNA和基因组测序已经在NCBI数据库中产生了数千个参考序列。虽然cDNA序列和基因组测序提供了重要的结构信息，但如果要继续研究基因功能或蛋白质相互作用，获得全长克隆仍然是关键。公共数据库中的许多汇编基因序列没有可用于支持进一步研究的物理全长克隆。OriGene Technologies，Inc.已经开发了超过20个最先进的cDNA文库（包括6个小鼠cDNA文库），即快速筛选阵列cDNA文库面板，以支持高通量全长cDNA克隆。迄今为止，我们已经发布了24，000个独特的人类全长cDNA克隆，这是业界最大的，用于功能基因组学研究。由于人类和小鼠之间的遗传可塑性以及生物学和病理学相似性，小鼠已成为基础哺乳动物生物学和人类疾病的主要模式生物。许多小鼠实验操作需要一组小鼠全长cDNA克隆。该项目第一阶段的目标是在我们的RapidScreen cDNA文库中分离和测序50，000个富含大插入片段（>4 kb）的小鼠cDNA克隆。这将包括与现有的小鼠克隆库（如MGC和RIKEN）进行比较时，新全长cDNA克隆的鉴定和确认。该项目的第二阶段将侧重于通过测序多达50万个克隆来大规模测序和鉴定新的全长cDNA克隆。我们的最终目标是产生至少4000-5000个不同的小鼠全长cDNA克隆（特别是ORF>3 kb的cDNA），这些克隆目前不是MGC收集的一部分。我们的技术平台已被证明是可靠和创新的，该项目的成果将肯定促进当前和未来的小鼠功能基因组学研究。

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.