CRISPR-clear imaging of melanin-rich B16-derived solid tumors.
Commun Biol
; 6(1): 370, 2023 04 04.
Article
in En
| MEDLINE
| ID: mdl-37016073
ABSTRACT
Tissue clearing combined with deep imaging has emerged as a powerful technology to expand classical histological techniques. Current techniques have been optimized for imaging sparsely pigmented organs such as the mammalian brain. In contrast, melanin-rich pigmented tissue, of great interest in the investigation of melanomas, remains challenging. To address this challenge, we have developed a CRISPR-based gene editing approach that is easily incorporated into existing tissue-clearing workflows such the PACT clearing method. We term this method CRISPR-Clear. We demonstrate its applicability to highly melanin-rich B16-derived solid tumors, including one made transgenic for HER2, constituting one of very few syngeneic mouse tumors that can be used in immunocompetent models. We demonstrate the utility in detailed tumor characterization by staining for targeting antibodies and nanoparticles, as well as expressed fluorescent proteins. With CRISPR-Clear we have unprecedented access to optical interrogation in considerable portions of intact melanoma tissue for stained surface markers, expressed fluorescent proteins, of subcellular compartments, and of the vasculature.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Melanins
/
Melanoma
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
Commun Biol
Year:
2023
Document type:
Article
Affiliation country:
Switzerland