Genome Editing of Human Primary Keratinocytes by CRISPR/Cas9 Reveals an Essential Role of the NLRP1 Inflammasome in UVB Sensing
Open accessAbstract
By forming a protective barrier, epidermal keratinocytes represent the first line of defense against environmental insults. UVB radiation of the sun is a major challenge for the skin and can induce inflammation, aging, and eventually skin cancer. UVB induces an immune response in human keratinocytes resulting in activation and secretion of the proinflammatory cytokines proIL-1β and -18. This is mediated by an assembly of protein complexes, termed inflammasomes. However, the mechanisms underlying sensing of UVB by keratinocytes, and particularly the types of inflammasomes required for cytokine secretion, are a matter of debate. To address these questions, we established a protocol that allows the generation of CRISPR/Cas9-targeted human primary keratinocytes. Our experiments showed an essential role of the NLRP1 rather than the NLRP3 inflammasome in UVB sensing and subsequent IL-1β and -18 secretion by keratinocytes. Moreover, NLRP1 but not NLRP3 was required for inflammasome activation in response to nigericin, a potassium ionophore and well-established NLRP3 activator in immune cells. Because the CRISPR/Cas9-targeted cells retained their full differentiation capacity, genome editing of human primary keratinocytes might be useful for numerous research and medical applications.
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Antibiotic resistance-based selection methods for screening recipient cells result in low efficiency in HKs due to their short lifespan.
“Additionally, the antibiotic resistance–based selection methods associated with the screening of recipient cells results in low efficiency considering the short lifespan of HKs (Fenini et al., 2018).”
From CRISPR/Cas9 Ribonucleoprotein Nucleofection for Genome Editing in Primary Human Keratinocytes: Knockouts, Deletions, and Homology‐Directed Repair Mutagenesis (2024) · cites this paper as Fenini et al., 2018
In “Establishment of a protocol for stable genetic modification of keratinocytes” (page unavailable), the paper says antibiotic selection in co-culture enabled efficient modification of human primary keratinocytes, contrary to the claimed low efficiency. It notes that these cells stop proliferating after a few passages under certain culture conditions and that prolonged puromycin selection harms survival, but does not attribute low selection efficiency to their short lifespan. (LLM, conf=0.86)
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- Authors
- Fenini, G., Grossi, S., Contassot, E., Biedermann, T., Reichmann, E., French, L. E., & Beer, H.-D.
- Affiliations
- University Hospital Zurich (CH)
- University of Zurich (CH)
- University Children's Hospital Zurich (CH)
- Corresponding author
- Beer, Hans-Dietmar
- Venue
- Journal of Investigative Dermatology
- Publisher
- Elsevier BV
- Published
- 2018-12
- Volume
- 138
- Issue
- 12
- Pages
- 2644-2652
- ISSN
- 0022-202X (ISSN-L)
- OpenAlex
- W2886580824
- Cited by
- 125
- References
- 51
- Citations by year
- 2026: 7 · 2025: 15 · 2024: 23 · 2023: 19 · 2022: 17 · 2021: 11 · 2020: 22 · 2019: 8 · 2018: 2 · 2014: 1
- FWCI
- 3.82
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Journal of Investigative Dermatology (publishedVersion)
- PubMed (publishedVersion)
- Zurich Open Repository and Archive (University of Zurich) (acceptedVersion, other-oa)
- Zurich Open Repository and Archive (University of Zurich) (submittedVersion, other-oa)
- Zurich Open Repository and Archive (University of Zurich)
- Funders
- OPO-Stiftung
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (310030-156384, 31003A-120400)
- Promedica Stiftung
- Julius Müller Stiftung
- Kurt und Senta Herrmann Stiftung
- Wilhelm Sander-Stiftung
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Health Sciences › Medicine › Dermatology › Skin Protection and Aging
- Keywords
- NLRP1 inflammasome, NLRP3 inflammasome, keratinocytes, IL-1β secretion, inflammatory cytokines, nigericin, potassium ionophore, inflammasome activation, skin inflammation, CRISPR/Cas9 genome editing, skin barrier
- MeSH terms
- NLR Proteins, NLR Family, Pyrin Domain-Containing 3 Protein, Gene Editing, CRISPR-Associated Protein 9, Cells, Cultured, Humans, Nigericin, Radiodermatitis, Ultraviolet Rays, Keratinocytes, Interleukin-18, Adaptor Proteins, Signal Transducing, Apoptosis Regulatory Proteins, Interleukin-1beta, Inflammasomes, Primary Cell Culture, CRISPR-Cas Systems
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