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- W3198131653 abstract "Abstract In eukaryotic nuclei, chromatin loops mediated through cohesin are critical structures that regulate gene expression and DNA replication. Here, we demonstrate a new method to see endogenous genomic loci using synthetic zinc‐finger proteins harboring repeat epitope tags (ZF probes) for signal amplification via binding of tag‐specific intracellular antibodies, or frankenbodies, fused with fluorescent proteins. We achieve this in two steps: First, we develop an anti‐FLAG frankenbody that can bind FLAG‐tagged proteins in diverse live‐cell environments. The anti‐FLAG frankenbody complements the anti‐HA frankenbody, enabling two‐color signal amplification from FLAG‐ and HA‐tagged proteins. Second, we develop a pair of cell‐permeable ZF probes that specifically bind two endogenous chromatin loci predicted to be involved in chromatin looping. By coupling our anti‐FLAG and anti‐HA frankenbodies with FLAG‐ and HA‐tagged ZF probes, we simultaneously see the dynamics of the two loci in single living cells. This shows a close association between the two loci in the majority of cells, but the loci markedly separate from the triggered degradation of the cohesin subunit RAD21. Our ability to image two endogenous genomic loci simultaneously in single living cells provides a proof of principle that ZF probes coupled with frankenbodies are useful new tools for exploring genome dynamics in multiple colors." @default.
- W3198131653 created "2021-09-13" @default.
- W3198131653 creator A5020161951 @default.
- W3198131653 creator A5021449949 @default.
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- W3198131653 creator A5052892455 @default.
- W3198131653 creator A5054136956 @default.
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- W3198131653 creator A5068412904 @default.
- W3198131653 creator A5083891802 @default.
- W3198131653 creator A5085662599 @default.
- W3198131653 date "2021-09-20" @default.
- W3198131653 modified "2023-10-15" @default.
- W3198131653 title "Visualizing looping of two endogenous genomic loci using synthetic zinc‐finger proteins with anti‐FLAG and anti‐HA frankenbodies in living cells" @default.
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