Journal
GENOME BIOLOGY
Volume 19, Issue -, Pages -Publisher
BMC
DOI: 10.1186/s13059-018-1413-5
Keywords
Genome labeling; CRISPR/dCas9; Bimolecular fluorescence complementation (BIFC)
Funding
- Beijing municipal government [2017A02]
- National Thousand Young Talents Program of China
- National Natural Science Foundation of China [81572795, 81773304]
Ask authors/readers for more resources
CRISPR/dCas9 binds precisely to defined genomic sequences through targeting of guide RNA (gRNA) sequences. In vivo imaging of genomic loci can be achieved by recruiting fluorescent proteins using either dCas9 or gRNA. We thoroughly validate and compare the effectiveness and specificity of several dCas9/gRNA genome labeling systems. Surprisingly, we discover that in the gRNA-labeling strategies, accumulation of tagged gRNA transcripts leads to non-specific labeling foci. Furthermore, we develop novel bimolecular fluorescence complementation (BIFC) methods that combine the advantages of both dCas9-labeling and gRNA-labeling strategies. The BIFC-dCas9/gRNA methods demonstrate high signal-to-noise ratios and have no non-specific foci.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available