4. Direct capture PerturbSeq Perturb-Seq、CRISP-Seq和CROP-Seq等方法已经可以在单细胞的基础上分析目的基因、调控网络与表型之间的关系,但这几种方法仍然有缺陷: 1) 在Perturb-Seq和CRISP-Seq中,需要GBC或UGI标记sgRNA,才能检测出对应的sgRNA。但是在测序和生信分析等过程中存在误差,GBC或UGI与sgRNA配对有一定概率...
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...
CRISPLD2, which encodes a secreted protein previously implicated in lung development and endotoxin regulation, was found to have SNPs that were moderately associated with inhaled corticosteroid resistance and bronchodilator response among asthma patients in two previously conducted genome-wide association ...
RNA-Seq Transcriptome Profiling Identifies CRISPLD2 as a Glucocorticoid Responsive Gene that Modulat,程序员大本营,技术文章内容聚合第一站。
Direct capture PerturbSeq的两种建库测序方法与之前Perturb-Seq、CRISP-Seq和CROP-Seq等方法的检测效果一样好,并且一个慢病毒载体可以装载多个U6-sgRNA,极大地提高了单细胞扰动实验的效率。 四、小结 CRISPR 文库筛选技术具有高通量、精确筛选靶基因等特点,大大简化我们研究的实验流程和节省时间成本。在结合单细胞测序后...
Direct capture PerturbSeq的两种建库测序方法与之前Perturb-Seq、CRISP-Seq和CROP-Seq等方法的检测效果一样好,并且一个慢病毒载体可以装载多个U6-sgRNA,极大地提高了单细胞扰动实验的效率。 四、小结 CRISPR 文库筛选技术具有高通量、精确筛选靶基因等特点,大大简化我们研究的实验流程和节省时间成本。在结合单细胞测序后...
Direct capture PerturbSeq的两种建库测序方法与之前Perturb-Seq、CRISP-Seq和CROP-Seq等方法的检测效果一样好,并且一个慢病毒载体可以装载多个U6-sgRNA,极大地提高了单细胞扰动实验的效率。 四、小结 CRISPR 文库筛选技术具有高通量、精确筛选靶基因等特点,大大简化我们研究的实验流程和节省时间成本。在结合单细胞测序后...
CRISP-SEQ, AN INTEGRATED METHOD FOR MASSIVELY PARALLEL SINGLE CELL RNA-SEQ AND CRISPR POOLED SCREENSAn expression construct is disclosed which comprisesIdo AMITDiego JAITINDavid LARA-ASTIASOAssaf WEINERIdo YOFE