Country | Type | Number | Dated | Case |
United States Of America | Published Application | 20180002736 | 01/04/2018 | 2015-090 |
European Patent Office | Published Application | 3250689 | 12/06/2017 | 2015-090 |
Patent Cooperation Treaty | Published Application | WO2016123243 | 08/04/2016 | 2015-090 |
Genome
editing using CRISPR/Cas9 has enabled rapid and accessible alteration of
specific genomic loci in many organisms. A flexible means to target nucleic
acids would allow alteration and imaging of endogenous RNA transcripts, for
example, analogous to CRISPR/Cas-based genomic tools, but most nucleic acid
targeting methods rely on incorporation of exogenous tags.
UC Berkeley
researchers discovered compositions and methods for labeling a single stranded
target nucleic acid with the use of a Cas9 protein; a Cas9 guide RNA; and a quenched
PAMmer (a single stranded oligonucleotide having a protospacer adjacent motif
(PAM) sequence. The PAMmer also contains
a detectable label and a quencher moiety that quenches the detectable label. Cas9 cleavage of the PAMmer is predicted on
complete Cas9 sgRNA: target complementarity and thus is highly specific. The inventors have used the methods and compositions
to track RNA in living cells in a programmable manner without genetically
encoded tags.
Imaging, Cas9