CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes
Abstract Uncovering gene-phenotype relationships can be enabled by precise gene modulation in human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs) and follow up phenotyping using scalable all-optical electrophysiology platforms. Such efforts towards human functional genomics can be...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-12-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-023-05627-y |
| _version_ | 1797397836976881664 |
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| author | Julie L. Han Yuli W. Heinson Christianne J. Chua Wei Liu Emilia Entcheva |
| author_facet | Julie L. Han Yuli W. Heinson Christianne J. Chua Wei Liu Emilia Entcheva |
| author_sort | Julie L. Han |
| collection | DOAJ |
| description | Abstract Uncovering gene-phenotype relationships can be enabled by precise gene modulation in human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs) and follow up phenotyping using scalable all-optical electrophysiology platforms. Such efforts towards human functional genomics can be aided by recent CRISPR-derived technologies for reversible gene inhibition or activation (CRISPRi/a). We set out to characterize the performance of CRISPRi in post-differentiated iPSC-CMs, targeting key cardiac ion channel genes, KCNH2, KCNJ2, and GJA1, and providing a multiparametric quantification of the effects on cardiac repolarization, stability of the resting membrane potential and conduction properties using all-optical tools. More potent CRISPRi effectors, e.g., Zim3, and optimized viral delivery led to improved performance on par with the use of CRISPRi iPSC lines. Confirmed mild yet specific phenotype changes when CRISPRi is deployed in non-dividing differentiated heart cells is an important step towards more holistic pre-clinical cardiotoxicity testing and for future therapeutic use in vivo. |
| first_indexed | 2024-03-09T01:15:57Z |
| format | Article |
| id | doaj.art-c1567f50a3794f879ded230e7ed84f5d |
| institution | Directory Open Access Journal |
| issn | 2399-3642 |
| language | English |
| last_indexed | 2024-03-09T01:15:57Z |
| publishDate | 2023-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj.art-c1567f50a3794f879ded230e7ed84f5d2023-12-10T12:28:28ZengNature PortfolioCommunications Biology2399-36422023-12-016111610.1038/s42003-023-05627-yCRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytesJulie L. Han0Yuli W. Heinson1Christianne J. Chua2Wei Liu3Emilia Entcheva4Department of Biomedical Engineering, The George Washington UniversityDepartment of Biomedical Engineering, The George Washington UniversityDepartment of Biomedical Engineering, The George Washington UniversityDepartment of Biomedical Engineering, The George Washington UniversityDepartment of Biomedical Engineering, The George Washington UniversityAbstract Uncovering gene-phenotype relationships can be enabled by precise gene modulation in human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs) and follow up phenotyping using scalable all-optical electrophysiology platforms. Such efforts towards human functional genomics can be aided by recent CRISPR-derived technologies for reversible gene inhibition or activation (CRISPRi/a). We set out to characterize the performance of CRISPRi in post-differentiated iPSC-CMs, targeting key cardiac ion channel genes, KCNH2, KCNJ2, and GJA1, and providing a multiparametric quantification of the effects on cardiac repolarization, stability of the resting membrane potential and conduction properties using all-optical tools. More potent CRISPRi effectors, e.g., Zim3, and optimized viral delivery led to improved performance on par with the use of CRISPRi iPSC lines. Confirmed mild yet specific phenotype changes when CRISPRi is deployed in non-dividing differentiated heart cells is an important step towards more holistic pre-clinical cardiotoxicity testing and for future therapeutic use in vivo.https://doi.org/10.1038/s42003-023-05627-y |
| spellingShingle | Julie L. Han Yuli W. Heinson Christianne J. Chua Wei Liu Emilia Entcheva CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes Communications Biology |
| title | CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes |
| title_full | CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes |
| title_fullStr | CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes |
| title_full_unstemmed | CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes |
| title_short | CRISPRi gene modulation and all-optical electrophysiology in post-differentiated human iPSC-cardiomyocytes |
| title_sort | crispri gene modulation and all optical electrophysiology in post differentiated human ipsc cardiomyocytes |
| url | https://doi.org/10.1038/s42003-023-05627-y |
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