Spatial CRISPR screens in tumors - Nature Methods
2022年5月11日 · To this end, they introduced Perturb-map, which combines multiplex imaging and spatial transcriptomics to resolve the roles of perturbed genes in lung tumors by profiling the molecular state of...
Massively parallel phenotyping of coding variants in cancer with ...
2022年1月20日 · Here, we modified Perturb-seq for pooled genetic screens with single-cell RNA-sequencing (scRNA-seq) readout 37,38 to phenotype coding variants using single-cell expression-based variant...
Massively parallel phenotyping of coding variants in cancer with ...
2022年1月22日 · We measured the impact of 200 TP53 and KRAS variants on RNA profiles in over 300,000 single lung cancer cells, and used the profiles to categorize variants into phenotypic subsets to distinguish gain-of-function, loss-of-function and dominant negative variants, which we validated by comparison with orthogonal assays.
Spatial CRISPR genomics identifies regulators of the tumor ...
2022年3月31日 · We applied Perturb-map to knock out dozens of genes in parallel in a mouse model of lung cancer and simultaneously assessed how each knockout influenced tumor growth, histopathology, and immune composition. Moreover, we paired Perturb-map and spatial transcriptomics for unbiased analysis of CRISPR-edited tumors.
Massively Parallel CRISPR‐Based Genetic Perturbation Screening at ...
The pioneering techniques (Perturb-seq, CRISP-seq, Mosaic-seq) track sgRNA identity by inserting unique guide barcodes (GBC) into sgRNA-encoding plasmids. The endogenous mRNA and GBC transcripts are ligated by the same cell barcode (CBC) during reverse transcription (RT) within the built-in microchambers of single-cell platforms.
PerturbDB for unraveling gene functions and regulatory networks
2024年9月11日 · Perturb-Seq, an innovative and powerful technique, combines CRISPR (clustered regularly interspaced short palindromic repeats)-based screens with single-cell RNA sequencing (scRNA-Seq) readouts for high-content phenotypic screens to comprehensively map the transcriptional effects of genetic perturbations genome-wide . This technique is highly ...
Targeted Perturb-seq enables genome-scale genetic screens in …
2020年6月1日 · TAP-seq permits routine analysis of 1,000s of CRISPR-mediated perturbations within a single experiment, detects weak effects and lowly expressed genes, and decreases sequencing requirements up to 50-fold. We apply TAP-seq to generate perturbation-based enhancer-target gene maps for 1,778 enhancers within 2.5% of the human genome.
王炸组合！Perturb-Seq：CRISPR+single cell - 知乎 - 知乎专栏
2022年7月25日 · 通过开发用于体内应用的Perturb-Seq，我们可以开始以更有效的方式在动物模型中筛选所有这些基因，从而使我们能够从机理上理解这些基因突变是如 展开阅读全文
Perturb-Seq: Dissecting Molecular Circuits with Scalable
2016年12月15日 · We demonstrate Perturb-seq by analyzing 200,000 cells in immune cells and cell lines, focusing on transcription factors regulating the response of dendritic cells to lipopolysaccharide (LPS). Perturb-seq accurately identifies individual gene targets, gene signatures, and cell states affected by individual perturbations and their genetic ...
Systematic functional interrogation of SARS-CoV-2 host factors …
2023年10月6日 · Here we use Perturb-seq, combining genetic perturbations with a single-cell readout, to investigate how inactivation of host factors changes the course of SARS-CoV-2 infection and the host response in human lung epithelial cells.