Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic imaging systems predominantly rely on fluorescent protein reporters, which lack the optical properties essential for sensitive dynamic imaging. Here, we modified the CRISPR single-guide RNA (sgRNA) to carry two distinct molecular beacons (MBs) that can undergo fluorescence resonance energy transfer (FRET) and demonstrated that the resulting system, CRISPR/dual-FRET MB, enables dynamic imaging of non-repetitive genomic loci with only three unique sgRNAs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6847002 | PMC |
| http://dx.doi.org/10.1093/nar/gkz752 | DOI Listing |
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Article Synopsis
- Developed CRISPRdelight, a DNA-labeling system using dCas12a, to improve imaging of non-repetitive genomic loci and understand gene regulation.
- Achieved effective imaging of 12 non-repetitive loci across different cell lines, revealing the unique behaviors of specific genes like CCAT1 and HSPH1 in response to transcriptional activity and stress.
- Introduced multiplex imaging capabilities with CRISPR-dCas12a and RNA aptamers, allowing for simultaneous tracking of multiple satellite DNA loci and their interactions with nucleolus-associated domains.
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