Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes
Summary: Modulation of transcription, either synthetic activation or repression, via dCas9-fusion proteins is a relatively new methodology with the potential to facilitate high-throughput up- or downregulation studies of gene function. Genetic studies of neurodevelopmental disorders have identified...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2017-08-01
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| Series: | Stem Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213671117302758 |
| _version_ | 1818838068293009408 |
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| author | Seok-Man Ho Brigham J. Hartley Erin Flaherty Prashanth Rajarajan Rawan Abdelaal Ifeanyi Obiorah Natalie Barretto Hamza Muhammad Hemali P. Phatnani Schahram Akbarian Kristen J. Brennand |
| author_facet | Seok-Man Ho Brigham J. Hartley Erin Flaherty Prashanth Rajarajan Rawan Abdelaal Ifeanyi Obiorah Natalie Barretto Hamza Muhammad Hemali P. Phatnani Schahram Akbarian Kristen J. Brennand |
| author_sort | Seok-Man Ho |
| collection | DOAJ |
| description | Summary: Modulation of transcription, either synthetic activation or repression, via dCas9-fusion proteins is a relatively new methodology with the potential to facilitate high-throughput up- or downregulation studies of gene function. Genetic studies of neurodevelopmental disorders have identified a growing list of risk variants, including both common single-nucleotide variants and rare copy-number variations, many of which are associated with genes having limited functional annotations. By applying a CRISPR-mediated gene-activation/repression platform to populations of human-induced pluripotent stem cell-derived neural progenitor cells, neurons, and astrocytes, we demonstrate that it is possible to manipulate endogenous expression levels of candidate neuropsychiatric risk genes across these three cell types. Although proof-of-concept studies using catalytically inactive Cas9-fusion proteins to modulate transcription have been reported, here we present a detailed survey of the reproducibility of gRNA positional effects across a variety of neurodevelopmental disorder-relevant risk genes, donors, neural cell types, and dCas9 effectors. : Brennand and colleagues report a survey of the reproducibility of CRISPR-mediated transcriptional modulation across varied neurodevelopmental disorder risk genes, donors, neural cell types, and dCas9 effectors. We report a number of practical limitations that must be considered when designing hiPSC-based studies using this promising new tool. Keywords: CRISPR, human-induced pluripotent stem cell, neural progenitor cell, transcriptional modulation, dCas9-VP64, dCas9-VPR, dCas9-KRAB |
| first_indexed | 2024-12-19T03:32:31Z |
| format | Article |
| id | doaj.art-2772a37d93fb404291498f59170646ce |
| institution | Directory Open Access Journal |
| issn | 2213-6711 |
| language | English |
| last_indexed | 2024-12-19T03:32:31Z |
| publishDate | 2017-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Reports |
| spelling | doaj.art-2772a37d93fb404291498f59170646ce2022-12-21T20:37:27ZengElsevierStem Cell Reports2213-67112017-08-0192615628Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and AstrocytesSeok-Man Ho0Brigham J. Hartley1Erin Flaherty2Prashanth Rajarajan3Rawan Abdelaal4Ifeanyi Obiorah5Natalie Barretto6Hamza Muhammad7Hemali P. Phatnani8Schahram Akbarian9Kristen J. Brennand10Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USACenter for Genomics of Neurodegenerative Disease, New York Genome Center, New York, NY 10013, USA; Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, New York, NY 10032, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USACenter for Genomics of Neurodegenerative Disease, New York Genome Center, New York, NY 10013, USA; Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, New York, NY 10032, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Genetics and Genomics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Corresponding authorSummary: Modulation of transcription, either synthetic activation or repression, via dCas9-fusion proteins is a relatively new methodology with the potential to facilitate high-throughput up- or downregulation studies of gene function. Genetic studies of neurodevelopmental disorders have identified a growing list of risk variants, including both common single-nucleotide variants and rare copy-number variations, many of which are associated with genes having limited functional annotations. By applying a CRISPR-mediated gene-activation/repression platform to populations of human-induced pluripotent stem cell-derived neural progenitor cells, neurons, and astrocytes, we demonstrate that it is possible to manipulate endogenous expression levels of candidate neuropsychiatric risk genes across these three cell types. Although proof-of-concept studies using catalytically inactive Cas9-fusion proteins to modulate transcription have been reported, here we present a detailed survey of the reproducibility of gRNA positional effects across a variety of neurodevelopmental disorder-relevant risk genes, donors, neural cell types, and dCas9 effectors. : Brennand and colleagues report a survey of the reproducibility of CRISPR-mediated transcriptional modulation across varied neurodevelopmental disorder risk genes, donors, neural cell types, and dCas9 effectors. We report a number of practical limitations that must be considered when designing hiPSC-based studies using this promising new tool. Keywords: CRISPR, human-induced pluripotent stem cell, neural progenitor cell, transcriptional modulation, dCas9-VP64, dCas9-VPR, dCas9-KRABhttp://www.sciencedirect.com/science/article/pii/S2213671117302758 |
| spellingShingle | Seok-Man Ho Brigham J. Hartley Erin Flaherty Prashanth Rajarajan Rawan Abdelaal Ifeanyi Obiorah Natalie Barretto Hamza Muhammad Hemali P. Phatnani Schahram Akbarian Kristen J. Brennand Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes Stem Cell Reports |
| title | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes |
| title_full | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes |
| title_fullStr | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes |
| title_full_unstemmed | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes |
| title_short | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes |
| title_sort | evaluating synthetic activation and repression of neuropsychiatric related genes in hipsc derived npcs neurons and astrocytes |
| url | http://www.sciencedirect.com/science/article/pii/S2213671117302758 |
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