3)CROP-Seq是将U6-sgRNA表达盒大片段添加至Puro抗性基因后3′LTR区,这就限制了不能添加多个sgRNA。 Direct capture PerturbSeq则很好地解决了这些问题。Direct capture PerturbSeq是基于10×Gebomics公司的5′端和3′端的建库方法,开发了5′端和3′端两种检测sgRNA方案[5]。在5′端的检测方案中,针对sgRNA后面的...
如今,多亏两种高度类似的技术:一种技术是由美国布罗德研究所的Aviv Regev和同事们开发出的,该技术被称作Perturb-Seq[1];另一种技术是由以色列魏兹曼研究所的Ido Amit和同事们开发出的,该技术被称作CRISP-Seq[2]。在一项实验中,利用Perturb-Seq、CRISP-Seq或将这两种技术结合在一起研究众多基因扰动是可能的[3]。
CRISP-SEQ, AN INTEGRATED METHOD FOR MASSIVELY PARALLEL SINGLE CELL RNA-SEQ AND CRISPR POOLED SCREENSAn expression construct is disclosed which comprises: (i) a DNA sequence which encodes at least one guide RNA (g RNA) operatively linked to a transcriptional regulatory sequence so as to allow ...
在一项实验中,利用Perturb-Seq、CRISP-Seq或将这两种技术结合在一起研究众多基因扰动是可能的[3]。 Perturb-Seq和CRISP-Seq背后的工作原理是给单个基因扰动和受到影响的细胞添加条形码以至于利用测序能够鉴定出这两者。简单地说,靶向感兴趣基因的添加独特条形码的CRISPR向导RNA文库被导入一个细胞群体中。随后利用添加独特...
(homopolymer length or microsatellite length) FLANKSEQ: This represents the reference haplotype sequence (length spans the homopolymer or microsatellite tract) with 10 bases either side of the variant position (10bases_upstream:reference_haplotype_sequence:10bases_downstream). This is useful to eyeball ...
RNA-seq分析表明,CRISPR干扰(CRISPR interference, CRISPRi)介导的转录抑制是高度特异性的。研究结果表明,CRISPR系统可以作为一个模块化和灵活的DNA结合平台,用于将蛋白质募集到目标DNA序列,并揭示了CRISPRi作为精确调节真核细胞中基因表达的通用工具的潜力。
The V6 region of the 16S ribosomal RNA gene was amplified and sequenced using the Illumina HiSeq 2000 platform. Filtered culture supernatants from Lactobacillus crispatus, Lactobacillus iners, and Gardnerella vaginalis were also assayed for E. coli inhibitory activity.Sixteen samples (10 with high ...
Using RNA-Seq, a high-throughput sequencing method, we characterized transcriptomic changes in four primary human ASM cell lines that were treated with dexamethasone—a potent synthetic glucocorticoid (1 µM for 18 hours). Based on a Benjamini-Hochberg corrected p-value <0.05, we identified 316...
RNA-seq分析表明,CRISPR干扰介导的转录抑制是高度特异性的。研究结果表明,CRISPR系统可以作为一个模块化和灵活的DNA结合平台,用于将蛋白质募集到目标DNA序列,并揭示了CRISPRi作为精确调节真核细胞中基因表达的通用工具的潜力。 自此,基因编辑的新篇章正式拉开帷幕! PART 3 CRISPR剪出发展蓝图 CRISPR-Cas9 技术具有许多...
(<20% inhibition) anti-E. coli activity. The V6 region of the 16S ribosomal RNA gene was amplified and sequenced using the Illumina HiSeq 2000 platform. Filtered culture supernatants from Lactobacillus crispatus, Lactobacillus iners, and Gardnerella vaginalis were also assayed for E. coli ...