然后作者用CRISPRi-seq,对目前肺炎链球菌领域里常用的小鼠肺炎模型进行了研究,发现该模型中细菌侵染过程存在着显著的瓶颈效应:起始构建侵染模型时用的菌量是108CFU,侵染过程中有的小鼠体内的菌量可以降到25个,然而这仅仅25个存活下来的菌便可以...
一、Highlight——CRISPRi–TnSeq工具 使用CRISPRi–TnSeq工具在全基因组范围内解释必需和非必需基因之间的合成和抑制关系,主要有以下五个步骤(以肺炎链球菌为例): (1)肺炎链球菌D39 CRISPRi菌株携带一个具有P3启动子的必需基因特异性sgRNA,和位于Plac启动子下游的dCas9。sgRNA与dCas9之间被LacI(带有PF6启动子)阻断。
(CRISPRi-seq). We demonstrate the use of the protocol inStreptococcus pneumoniae, an important human pathogen; however, the protocol can easily be adapted for use in other organisms. The protocol includes a pipeline for single-guide RNA library design, workflows for pooled CRISPRi library ...
To initially characterize pooled growth, the EGML strains were equally combined in a pooled inoculum (1X pool), allowed to grow at different rhamnose concentrations (Figure S7), and their relative abundance was determined by CRISPRi-seq. Non-inducing conditions (likely causing basal levels of ...
A CRISPRi-seq based whole genome screen was employed to identify the potential targets of azalomycin F4a, which revealed that peptidoglycan synthesis (PGS) was inhibited by azalomycin F4a. Furthermore, azalomycin F4a treatment could significantly impair S. aureus biofilm formation. Our research ...
Analyzing data from two large-scale CRISRPi screens in iPSCs with single-cell RNA-seq read-out. - claudiafeng123/crispri_scrnaseq_hipsci
CRISPRi细胞系 实验组/处理组的 ATAC-Seq 的测序。 基础生信报告 基与于 Pathcards 的基因注释与文献信息三级联转录调控预测。 本司不提供不确保: 基础生信报告达到SCI 20分以上Fig要求。 三级联转录调控轴心Axis预测,直接可写入SCI 5分以上论文结果。
Here we use CRISPRi–TnSeq, CRISPRi-mediated knockdown of essential genes alongside TnSeq-mediated knockout of non-essential genes, to map genome-wide interactions between essential and non-essential genes in. Transposon-mutant libraries constructed in 13 CRISPRi strains enabled screening of ~24,...
Although genetic predisposition has mainly been described in prostate cancer (PrCa), functional characterization of these risk loci remains a challenge.Methods: Low multiplicity of infection creates single lentiviral integrated cell population, which enable us to evaluate biological significance of steric ...
(ECs) identify causal genes and disease-relevant pathways at CAD GWAS loci.Methods: We applied CRISPRi-Perturb-seq to knock down expression of genes within 500 Kb of coronary artery disease GWAS loci (2,300 genes in total) and measure their transcriptomic effects using single-cell RNA-seq. ...