期刊
CYTOGENETIC AND GENOME RESEARCH
卷 160, 期 3, 页码 156-165出版社
KARGER
DOI: 10.1159/000506720
关键词
Cas9; CRISPR-FISH; FISH; Mouse; RGEN-ISL; Soybean; Super-resolution microscopy
资金
- DFG [Ho1779/28-1]
Visualizing the spatiotemporal organization of the genome will improve our understanding of how chromatin structure and function are intertwined. Here, we describe the further development of the RNA-guided endonuclease-in situ labeling (RGEN-ISL) method CRISPR-FISH. Using soybean and mouse chromosomes, we demonstrate that the treatment of conventionally fixed chromosomes (in ethanol or methanol:acetic acid) with 40 mM Tris-HCl (pH 9.0) for 30 minutes at 37 degrees C prior to CRISPR-FISH allows the application of this method for the detection of high-copy sequences. Wheat, rye, maize, andNicotiana benthamianawere used to confirm the applicability of the identified CRISPR-FISH conditions also in other species.
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