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Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...

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Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C

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在非编码基因组中的功能元素的高分辨率查询

Neville E Sanjana¹, Jason Wright², Kaijie Zheng²

¹Broad Institute of MIT and Harvard, 7 Cambridge Center, Cambridge, MA 02142, USA. McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA. nsanjana@nygenome.org zhang@broadinstitute.org.

Science (New York, N.Y.)

|October 7, 2016

概括

研究人员开发了CRISPR镜来识别影响黑色素瘤耐药性的非编码元素. 这些功能性非编码区域影响基因调节,可以作为新疗法的点.

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Screening for Functional Non-coding Genetic Variants Using Electrophoretic Mobility Shift Assay EMSA and DNA-affinity Precipitation Assay DAPA

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High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture 4C-seq

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Screening for Functional Non-coding Genetic Variants Using Electrophoretic Mobility Shift Assay EMSA and DNA-affinity Precipitation Assay DAPA

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科学领域:

基因组学
分子生物学
癌症研究

背景情况:

非编码基因组在基因调节和疾病发病过程中起着至关重要的作用.
目前用于识别和处理非编码元素的工具有限,阻碍了研究.
黑色素瘤对BRAF抑制剂的耐药性是一个重大的临床挑战.

研究的目的:

开发和应用CRISPR选方法,用于快速识别和功能性表征非编码基因组元素.
研究非编码区域在黑色素瘤中BRAF抑制剂耐药性的作用.
探索针对治疗策略的非编码元素的潜力.

主要方法:

使用集成的CRISPR屏幕,其中约有18,000个单指导RNA针对700多个围绕NF1,NF2和CUL3基因的基因.
对调节药物耐药性的功能重要特征进行分析.
在特定的CUL3位点进行工程突变,以评估对转录因子结合和表观遗传修饰的影响.

主要成果:

在黑色素瘤中显著调节BRAF抑制剂耐药性的非编码部位.
发现这些功能性非编码区域具有监管活动的预测特征.
证明CUL3位点非编码区域的工程突变改变了转录因子占用率和表观遗传特征,影响了基因调节和耐药性.

结论:

开发了新的CRISPR选工具,以有效地发现功能性非编码元素.
确立了特定非编码区域,基因调节和黑色素瘤化疗耐药性之间的联系.
隐含的非编码元素作为克服癌症耐药性的潜在治疗点.