Conditional Manipulation of Gene Function in Human Cells with Optimized Inducible shRNA Article Swipe
Alessandro Bertero
,
Loukia Yiangou
,
Stephanie E. Brown
,
Daniel Ortmann
,
Matthias Pawlowski
,
Ludovic Vallier
·
YOU?
·
· 2018
· Open Access
·
· DOI: https://doi.org/10.1002/cpsc.45
YOU?
·
· 2018
· Open Access
·
· DOI: https://doi.org/10.1002/cpsc.45
The difficulties involved in conditionally perturbing complex gene expression networks represent major challenges toward defining the mechanisms controlling human development, physiology, and disease. We developed an OPTimized inducible KnockDown (OPTiKD) platform that addresses the limitations of previous approaches by allowing streamlined, tightly‐controlled, and potent loss‐of‐function experiments for both single and multiple genes. The method relies on single‐step genetic engineering of the AAVS1 genomic safe harbor with an optimized tetracycline‐responsive cassette driving one or more inducible short hairpin RNAs (shRNAs). OPTiKD provides homogeneous, dose‐responsive, and reversible gene knockdown. When implemented in human pluripotent stem cells (hPSCs), the approach can be then applied to a broad range of hPSC‐derived mature cell lineages that include neurons, cardiomyocytes, and hepatocytes. Generation of OPTiKD hPSCs in commonly used culture conditions is simple (plasmid based), rapid (two weeks), and highly efficient (>95%). Overall, this method facilitates the functional annotation of the human genome in health and disease. © 2018 by John Wiley & Sons, Inc.
Related Topics
Concepts
Gene knockdown
Induced pluripotent stem cell
Biology
Small hairpin RNA
Computational biology
Gene
RNA interference
Function (biology)
Cell biology
Genetics
RNA
Embryonic stem cell
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1002/cpsc.45
- OA Status
- green
- Cited By
- 14
- References
- 82
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W2790258422
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W2790258422Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1002/cpsc.45Digital Object Identifier
- Title
-
Conditional Manipulation of Gene Function in Human Cells with Optimized Inducible shRNAWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2018Year of publication
- Publication date
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2018-02-01Full publication date if available
- Authors
-
Alessandro Bertero, Loukia Yiangou, Stephanie E. Brown, Daniel Ortmann, Matthias Pawlowski, Ludovic VallierList of authors in order
- Landing page
-
https://doi.org/10.1002/cpsc.45Publisher landing page
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
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https://hdl.handle.net/2318/1804550Direct OA link when available
- Concepts
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Gene knockdown, Induced pluripotent stem cell, Biology, Small hairpin RNA, Computational biology, Gene, RNA interference, Function (biology), Cell biology, Genetics, RNA, Embryonic stem cellTop concepts (fields/topics) attached by OpenAlex
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-
14Total citation count in OpenAlex
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2025: 1, 2024: 3, 2023: 2, 2022: 2, 2021: 2Per-year citation counts (last 5 years)
- References (count)
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82Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.simple | 127 |
| abstract_inverted_index.single | 49 |
| abstract_inverted_index.toward | 14 |
| abstract_inverted_index.applied | 101 |
| abstract_inverted_index.based), | 129 |
| abstract_inverted_index.complex | 7 |
| abstract_inverted_index.culture | 124 |
| abstract_inverted_index.driving | 71 |
| abstract_inverted_index.genetic | 58 |
| abstract_inverted_index.genomic | 63 |
| abstract_inverted_index.hairpin | 77 |
| abstract_inverted_index.include | 112 |
| abstract_inverted_index.weeks), | 132 |
| abstract_inverted_index.(OPTiKD) | 30 |
| abstract_inverted_index.(hPSCs), | 95 |
| abstract_inverted_index.(plasmid | 128 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.Overall, | 137 |
| abstract_inverted_index.allowing | 40 |
| abstract_inverted_index.approach | 97 |
| abstract_inverted_index.cassette | 70 |
| abstract_inverted_index.commonly | 122 |
| abstract_inverted_index.defining | 15 |
| abstract_inverted_index.disease. | 23, 151 |
| abstract_inverted_index.involved | 3 |
| abstract_inverted_index.lineages | 110 |
| abstract_inverted_index.multiple | 51 |
| abstract_inverted_index.networks | 10 |
| abstract_inverted_index.neurons, | 113 |
| abstract_inverted_index.platform | 31 |
| abstract_inverted_index.previous | 37 |
| abstract_inverted_index.provides | 81 |
| abstract_inverted_index.(shRNAs). | 79 |
| abstract_inverted_index.KnockDown | 29 |
| abstract_inverted_index.OPTimized | 27 |
| abstract_inverted_index.addresses | 33 |
| abstract_inverted_index.developed | 25 |
| abstract_inverted_index.efficient | 135 |
| abstract_inverted_index.inducible | 28, 75 |
| abstract_inverted_index.optimized | 68 |
| abstract_inverted_index.represent | 11 |
| abstract_inverted_index.(>95%). | 136 |
| abstract_inverted_index.Generation | 117 |
| abstract_inverted_index.annotation | 143 |
| abstract_inverted_index.approaches | 38 |
| abstract_inverted_index.challenges | 13 |
| abstract_inverted_index.conditions | 125 |
| abstract_inverted_index.expression | 9 |
| abstract_inverted_index.functional | 142 |
| abstract_inverted_index.knockdown. | 87 |
| abstract_inverted_index.mechanisms | 17 |
| abstract_inverted_index.perturbing | 6 |
| abstract_inverted_index.reversible | 85 |
| abstract_inverted_index.controlling | 18 |
| abstract_inverted_index.engineering | 59 |
| abstract_inverted_index.experiments | 46 |
| abstract_inverted_index.facilitates | 140 |
| abstract_inverted_index.implemented | 89 |
| abstract_inverted_index.limitations | 35 |
| abstract_inverted_index.physiology, | 21 |
| abstract_inverted_index.pluripotent | 92 |
| abstract_inverted_index.development, | 20 |
| abstract_inverted_index.difficulties | 2 |
| abstract_inverted_index.hepatocytes. | 116 |
| abstract_inverted_index.homogeneous, | 82 |
| abstract_inverted_index.streamlined, | 41 |
| abstract_inverted_index.conditionally | 5 |
| abstract_inverted_index.single‐step | 57 |
| abstract_inverted_index.hPSC‐derived | 107 |
| abstract_inverted_index.cardiomyocytes, | 114 |
| abstract_inverted_index.dose‐responsive, | 83 |
| abstract_inverted_index.loss‐of‐function | 45 |
| abstract_inverted_index.tightly‐controlled, | 42 |
| abstract_inverted_index.tetracycline‐responsive | 69 |
| cited_by_percentile_year.max | 98 |
| cited_by_percentile_year.min | 91 |
| countries_distinct_count | 2 |
| institutions_distinct_count | 6 |
| citation_normalized_percentile.value | 0.6544734 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |