Скачать или смотреть CRISPR-based functional genomics in iPSC-based models of brain disease │Dr. Martin Kampmann

Date: Thursday, 30 July 2020

Abstract:
Human genes associated with brain-related diseases are being discovered at an accelerating pace. A major challenge is the identification of the mechanisms through which these genes act, and of potential therapeutic strategies. To elucidate such mechanisms in human cells, we established a CRISPR-based platform for genetic screening in human iPSC-derived neurons, astrocytes and microglia. Our approach relies on CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa), in which a catalytically dead version of the bacterial Cas9 protein recruits transcriptional repressors or activators, respectively, to endogenous genes to control their expression, as directed by a small guide RNA (sgRNA). Complex libraries of sgRNAs enable us to conduct genome-wide or focused loss-of-function and gain-of-function screens. Such screens uncover molecular players for phenotypes based on survival, stress resistance, fluorescent phenotypes, high-content imaging and single-cell RNA-Seq. To uncover disease mechanisms and therapeutic targets, we are conducting genetic modifier screens for disease-relevant cellular phenotypes in patient-derived neurons and glia with familial mutations and isogenic controls. In a genome-wide screen, we have uncovered genes that modulate the formation of disease-associated aggregates of tau in neurons with a tauopathy-linked mutation (MAPT V337M). CRISPRi/a can also be used to model and functionally evaluate disease-associated changes in gene expression, such as those caused by eQTLs, haploinsufficiency, or disease states of brain cells. We will discuss an application to Alzheimer’s Disease-associated genes in microglia.

Bio:
Dr Kampmann is an Associate Professor in the UCSF Department of Biochemistry and Biophysics and the Institute for Neurodegenerative Diseases, and an Investigator at the Chan Zuckerberg Biohub. He received his BA in Biochemistry from Cambridge University and his PhD in Biophysics/Cell Biology from Rockefeller University. The goal of Dr Kampmann’s research is to elucidate cellular mechanisms of brain disease and to develop new therapeutic strategies. He co-developed the CRISPRi and CRISPRa screening technologies, and his lab has pioneered CRISPR-based functional genomics in cell types derived from induced pluripotent stem cells (iPSCs). A major focus is the investigation of neurodegenerative diseases in human iPSC-derived neurons, astrocytes, and microglia, and 3D assembloids/organoids. Dr Kampmann was named an NIH Director’s New Innovator, an Allen Distinguished Investigator, a Chan Zuckerberg Biohub Investigator, and he received the CZI Ben Barres Early Career Acceleration Award.

Moderator:
Mosab Ali Awadelkareem, Al-Neelain University, Sudan.

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