Module 1: Generating and Genome Editing iPS Cells
Learn basic approaches for generating, reprogramming, and genetically screening induced pluripotent stem cell-derived (iPSC) neurons and glia.
 Valentina Lo Sardo, PhD Scripps Research |
 Martin Kampmann, PhD University of California, San Francisco |
After reviewing Module 1, participants at all career stages should be able to:
- Explain the basics of reprogramming.
- Describe how to make a study design considering possible limitations and proper number of samples.
- Summarize how to perform a reprogramming experiment.
- Summarize how to validate and establish iPSC lines.
- Outline genome editing techniques and how to proceed to design and perform an experiment.
- Explain CRISPR-based approaches to probe gene function in iPSC-derived neurons and glia.
- Describe different types of large-scale genetic screens in iPSC-derived neurons and glia.
-
Module 1A: Generating and Genome Editing iPS CellsIn this presentation, Valentina Lo Sardo will introduce the generation of iPSCs from somatic cells and methods of genome editing. Specifically, Lo Sardo will:
- Introduce reprogramming, including different cell sources and methods for generating iPSCs.
- Describe experimental design and procedure to generate, validate, and maintain iPSCs from peripheral blood mononuclear cells (PBMCs), including general considerations on source of variability in iPSCs.
- Introduce genome editing techniques.
- Explain experimental design and procedure to perform genome editing using TALENs, including general considerations on study design and type of controls.
-
Module 1B: Functional Genomics in iPSC-Derived Neurons and GliaIn this presentation, Martin Kampmann will introduce functional genomics approaches in iPSC-derived neurons and glia. 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.
- Use of functional genomics to understand mechanisms underlying disease-linked genes and potential therapeutic strategies.
Module 1A: Generating and Genome Editing iPS Cells
In this presentation, Valentina Lo Sardo will introduce the generation of iPSCs from somatic cells and methods of genome editing. Specifically, Lo Sardo will:
- Introduce reprogramming, including different cell sources and methods for generating iPSCs.
- Describe experimental design and procedure to generate, validate, and maintain iPSCs from peripheral blood mononuclear cells (PBMCs), including general considerations on source of variability in iPSCs.
- Introduce genome editing techniques.
- Explain experimental design and procedure to perform genome editing using TALENs, including general considerations on study design and type of controls.
Learning Objectives
After watching this presentation, participants at all career stages should be able to:
- Explain the basics of reprogramming.
- Summarize how to make a study design considering possible limitations and proper number of samples.
- Outline how to perform a reprogramming experiment.
- Explain how to validate and establish iPSC lines.
- Describe genome editing techniques and how to design and perform an experiment.
- Describe how to design the proper study based on your interests.
Supporting Resources
Download this guide to access resources that supplement Lo Sardo’s presentation.
Speaker
Valentina Lo Sardo, PhD
Valentina Lo Sardo is a staff scientist in the department of neuroscience and Dorris Neuroscience Center at Scripps Research. Lo Sardo’s research focuses on genomic stability and integrity of induced pluripotent stem cells (iPSCs), and she is interested in using iPSCs and genome editing to study genomic loci associated with human diseases. She earned her MS and PhD in pharmaceutical biotechnologies from the University of Milan, in Italy, where she worked in the laboratory of Elena Cattaneo and contributed to deciphering the role of the huntingtin gene in neural development and its aberrant function in Huntington disease. Lo Sardo completed postdoctoral work with Kristin Baldwin at Scripps Research, investigating the impact of aging in iPSCs’ epigenetic and genomic stability and applying genome editing of large genomic risk haplotypes and patient-specific iPSCs to uncover the function of a mysterious gene desert genomic locus influencing susceptibility for cardiovascular disease.
Module 1B: Functional Genomics in iPSC-Derived Neurons and Glia
In this presentation, Martin Kampmann will introduce functional genomics approaches in iPSC-derived neurons and glia. 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, by FACS
- Single-cell RNA sequencing
- High-content imaging
- Use of functional genomics to understand mechanisms underlying disease-linked genes and potential therapeutic strategies.
Learning Objectives
After watching this presentation, participants at all career stages should be able to:
- Outline CRISPR-based approaches to probe gene function in iPSC-derived neurons and glia.
- Explain different types of large-scale genetic screens in iPSC-derived neurons and glia.
Supporting Resources
Download this guide to access resources that supplement Kampmann’s presentation.
Speaker
Martin Kampmann, PhD
Martin Kampmann is an assistant professor at the University of California, San Francisco Institute for Neurodegenerative Diseases and department of biochemistry and biophysics. Kampmann received his BA in biochemistry from Cambridge University, in the United Kingdom, and his PhD in biophysics and cell biology from The Rockefeller University. As a postdoctoral fellow at UCSF, he spearheaded the development of next-generation functional genomics approaches, including systematic genetic interaction maps and CRISPR-based gain- and loss-of-function screens. The goal of Kampmann’s research is to elucidate cellular mechanisms of human diseases and to develop new therapeutic strategies. For this purpose, his lab has pioneered CRISPR-based genetic screening technology in cell types derived from induced pluripotent stem cells (iPSCs) and is investigating neurodegenerative diseases in human iPSC-derived neurons, astrocytes, and microglia.
