CRISPRi and CRISPRa genetic screens in cells derived from human induced pluripotent stem cells (hiPSCs) can reveal mechanisms of disease-associated genes and of selective vulnerability of specific cell types. We use biochemistry, biophysics and cell biology to "zoom in" on individual nodes of the network and to reveal their mechanism of action.

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Martin KAMPMANN, Assistant Professor | Cited by 3,206 | of University of California, San Francisco, CA (UCSF) | Read 139 publications | Contact Martin KAMPMANN

Specifically, Kampmann will: Cover CRISPRi (interference) and CRISPRa (activation) control of gene expression. Describe how to implement CRISPRi/a in iPSC-derived cells. Introduce large-scale genetic screens based on survival, fluorescent readouts of cell function, single-cell RNA sequencing, and high-content imaging. Kampmann explains: "For CRISPRi, we target a transcriptional repressor domain (the KRAB domain) to the transcription start site of genes to repress their expression.

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Wang T, Yu H, Hughes NW, et al. Gene Essentiality Profiling Reveals Gene Networks and Synthetic Lethal Interactions with Oncogenic Ras. Cell 2017;168:890-903.e15. CRISPRi survival screen in human iPSC-derived glutamatergic neurons at Day 14 (standard medium). Experiment. Twelve 10-cm Matrigel-coated dishes were each seeded with 4*106 CRISPRi-iPSCs infected with virus for the H1 CRISPRi-v2 sgRNA library in N2 Pre-Differentiation Medium (day -3) and differentiated by doxycyclin-induced Ngn2 expression. CRISPRi and CRISPRa genetic screens in cells derived from human induced pluripotent stem cells (hiPSCs) can reveal mechanisms of disease-associated genes and of selective vulnerability of specific cell types. We use biochemistry, biophysics and cell biology to "zoom in" on individual nodes of the network and to reveal their mechanism of action.

Disclosures: Martin Kampmann has filed a patent application related to CRISPRi and CRISPRa screening (PCT/US15/40449) and serves on the Scientific Advisory Board of Engine Biosciences.

Neurodegenerative, neurodevelopmental and neuropsychiatric disorders are among the greatest public health challenges, as many lack disease-modifying treatments. A major reason for the absence of effective therapies is our limited understanding of the causative molecular and cellular mechanisms.

2020-03-04 · Analysis of CRISPRi screen results was carried out using custom code (MAGeCK-iNC) developed in the Kampmann lab, which was previously described 37 and is freely available at https://kampmannlab

Kampmann crispri

2021-04-19 CRISPRi survival screen in human iPSC-derived glutamatergic neurons at Day 14 (standard medium). Experiment. Twelve 10-cm Matrigel-coated dishes were each seeded with 4*106 CRISPRi-iPSCs infected with virus for the H1 CRISPRi-v2 sgRNA library in N2 Pre-Differentiation Medium (day -3) and differentiated by doxycyclin-induced Ngn2 expression.. On Day 14, dead (floating cells) were removed … As a result, unlike standard CRISPR-Cas9, Kampmann predicted, CRISPRi shouldn't be toxic to iPSCs or stem cell-derived neurons. In the new paper, Kampmann and his collaborators describe how they adapted CRISPRi for use in human iPSCs and iPSC-derived neurons, and found that it could target and interfere with genes without killing the cell -- a feat that had long eluded scientists. Martin Kampmann's profile, publications, research topics, and co-authors.

Kampmann crispri

A central goal of research for targeted cancer therapy, or precision oncology, is to reveal the intrinsic vulnerabilities of  CRISPRi and CRISPRa CRISPRi(nterference) and CRISPRa(ctivation) The application of the bacterial CRISPR/Cas9 system in mammalian cells has  20 Oct 2020 Interested in CRISPRi in iPSCs and derived cell types? Together with @ AllenInstitute & @WardLab_NIH, we generated a highly characterized,  Kampmann spearheaded the development of a functional genomics platform that makes it possible to robustly identify human genes relevant to a cellular process   15 Aug 2019 So Kampmann decided to tackle the toxicity problem. As a postdoc in the lab of UCSF Professor Jonathan Weissman, PhD, Kampmann co-  Gilbert, L. A. et al. Genome-scale CRISPR-mediated control of gene repression and activation. Cell159, 647–661 (2014). Kampmann,  21 Jul 2017 The rapid adoption of CRISPR technology has enabled biomedical Kampmann M, Horlbeck MA, Chen Y, Tsai JC, Bassik MC, Gilbert LA,  28 Jul 2020 One of them is CRISPR/Cas9 based genome and transcriptome editing.
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When CRISPRi finds the gene it’s seeking, it suppresses its activity without making any cuts. In CRISPRi, Kampmann and his colleagues figured out a way to disable the scissors protein, Cas9, and attach instead a protein that blocks normal gene activity. The end result is a modified CRISPR that dampens gene activity without editing the DNA itself. Neurodegenerative, neurodevelopmental and neuropsychiatric disorders are among the greatest public health challenges, as many lack disease-modifying treatments.

As a postdoc in the lab of UCSF Professor Jonathan Weissman, PhD, Kampmann co-invented a tool known as CRISPRi Genome-wide screening then utilizes cells stably expressing dCas9-KRAB (CRISPRi), photoactivatable fluorescent protein (PA-mCherry), and a lentiviral guide RNA (gRNA) pool. Cells are screened by using microscopy and classified by artificial intelligence (AI) algorithms, which precisely identify the genetically altered phenotype.
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Kampmann crispri





2021-04-08 · Focusing on lipid oxidation, Kampmann's group next employed CRISPRi screens to hunt for genes that influenced levels of ROS or peroxidized lipids. Here, they found many expected genes, such as those involved in electron transport or autophagy—pathways that can cause and curtail oxidative stress, respectively.

However, most previous CRISPR-based screens were conducted in cancer cell lines rather than healthy, differentiated cells. Here, we describe a CRISPR interference (CRISPRi)-based platform for gene … 2019-10-23 and CRISPRi screening platforms each have their advantages for specific applications (Kampmann, 2018; Rosenbluh et al., 2017) but generally yield similar results (Horlbeck et al., CRISPRi/a can also be used to model and functionally evaluate disease-associated changes in gene expression, Dr. Kampmann is an associate professor at the University of California, 2020-10-14 CRISPRi and CRISPRagenetic screening inhumaniPSC-derivedneurons.CRISPRi in iPSCs has previously been demon-strated [8], and our own unpublished results have recently established the fea-sibility of pooled CRISPRi-based screens in iPSC-derived neurons. Such neurons are powerful tools to study cellular mech-anisms of neurodegenerative diseases. iPSCs Previously, Kampmann lab developed a strategy to control specific knockdown of genes (CRISPRi) in human neurons (Tian et al., 2019), now they expand this toolkit to control specific gene activation (CRISPRa).