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工程改造CRISPR-Cas9 复合体 ：一种强有力的全基因组功能基因筛选方法An engineered CRISPR-Cas9 complex：a robust method for screening for functional gene in genome-scale ;;OUTLINE;Methods for elucidating gene function;;;;CRISPR Adaptive Immunity;;;Research;;;;;MS2? bacteriophage coat proteins grab other proteins p65、VP64、HSF1? activation domains recruits a distinct subset of transcription factors and chromatin remodelling complexes;12genes difficult to activate using dCas9–VP64 and individual sgRNA1.0 guides ;For inter-gene variability：the basal expression level of each gene largely determines the level of activation. ;For intragenic variability: targeting within the -200 bp to +1 bp window;;;Genome-scale gene activation screen;enrichment of specific sgRNAs after PLX-4720 treatment; individual sgRNA and cDNA overexpression validation;engineered CRISPR-Cas9 complex—— a powerful genetic perturbation technology!;Silvana Konermann, Mark D. Brigham, Alexandro E. Trevino, Julia Joung, Omar O. AbudayyehClea Barcena, Patrick D. Hsu, Naomi Habib, Jonathan S. Gootenberg, Hiroshi Nishimasu, Osamu Nureki Feng Zhang. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 14136(2014) Hsu, P. D., Lander, E. S. Zhang,F. Development and applications of CRISPR-Cas9 for genome engineering. Cell157,1262–1278 (2014) Nishimasu, H.et al. Crystal structure of Cas9 in complex with guide RNA and targetDNA.Cell156,935–949 (2014) Shalem, O.et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science343,84–87 (2014);Thanks for;阅读大事记