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Skarnes WC Rosen B West AP Koutsourakis M Bushell W Iyer V Mujica AO Thomas M Harrow J Cox T Jackson D Severin J Biggs P Fu J Nefedov M de Jong PJ Stewart AF Bradley A 《Nature》2011,474(7351):337-342
Gene targeting in embryonic stem cells has become the principal technology for manipulation of the mouse genome, offering unrivalled accuracy in allele design and access to conditional mutagenesis. To bring these advantages to the wider research community, large-scale mouse knockout programmes are producing a permanent resource of targeted mutations in all protein-coding genes. Here we report the establishment of a high-throughput gene-targeting pipeline for the generation of reporter-tagged, conditional alleles. Computational allele design, 96-well modular vector construction and high-efficiency gene-targeting strategies have been combined to mutate genes on an unprecedented scale. So far, more than 12,000 vectors and 9,000 conditional targeted alleles have been produced in highly germline-competent C57BL/6N embryonic stem cells. High-throughput genome engineering highlighted by this study is broadly applicable to rat and human stem cells and provides a foundation for future genome-wide efforts aimed at deciphering the function of all genes encoded by the mammalian genome. 相似文献
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Chapman MA Lawrence MS Keats JJ Cibulskis K Sougnez C Schinzel AC Harview CL Brunet JP Ahmann GJ Adli M Anderson KC Ardlie KG Auclair D Baker A Bergsagel PL Bernstein BE Drier Y Fonseca R Gabriel SB Hofmeister CC Jagannath S Jakubowiak AJ Krishnan A Levy J Liefeld T Lonial S Mahan S Mfuko B Monti S Perkins LM Onofrio R Pugh TJ Rajkumar SV Ramos AH Siegel DS Sivachenko A Stewart AK Trudel S Vij R Voet D Winckler W Zimmerman T Carpten J Trent J Hahn WC Garraway LA Meyerson M Lander ES Getz G 《Nature》2011,471(7339):467-472
Multiple myeloma is an incurable malignancy of plasma cells, and its pathogenesis is poorly understood. Here we report the massively parallel sequencing of 38 tumour genomes and their comparison to matched normal DNAs. Several new and unexpected oncogenic mechanisms were suggested by the pattern of somatic mutation across the data set. These include the mutation of genes involved in protein translation (seen in nearly half of the patients), genes involved in histone methylation, and genes involved in blood coagulation. In addition, a broader than anticipated role of NF-κB signalling was indicated by mutations in 11 members of the NF-κB pathway. Of potential immediate clinical relevance, activating mutations of the kinase BRAF were observed in 4% of patients, suggesting the evaluation of BRAF inhibitors in multiple myeloma clinical trials. These results indicate that cancer genome sequencing of large collections of samples will yield new insights into cancer not anticipated by existing knowledge. 相似文献
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