scholarly journals Visualizing looping of two endogenous genomic loci using synthetic zinc‐finger proteins with anti‐FLAG and anti‐HA frankenbodies in living cells

2021 ◽  
Author(s):  
Yang Liu ◽  
Ning Zhao ◽  
Masato T. Kanemaki ◽  
Yotaro Yamamoto ◽  
Yoshifusa Sadamura ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Proteins ◽  
Living Cells

scholarly journals Visualizing looping of two endogenous genomic loci using synthetic zinc-finger proteins with anti-FLAG and anti-HA frankenbodies in living cells

2021 ◽  
Author(s):  
Yang Liu ◽  
Ning Zhao ◽  
Masato T Kanemaki ◽  
Yotaro Yamamoto ◽  
Yoshifusa Sadamura ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Signal Amplification ◽  
Fluorescent Proteins ◽  
Close Association ◽  
Zinc Finger Proteins ◽  
Living Cells ◽  
Regulate Gene Expression ◽  
Cohesin Subunit ◽  
Single Living Cells ◽  
Single Living

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 visualize 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 visualize the dynamics of the two loci in single living cells. This reveals close association between the two loci in the majority of cells, but the loci markedly separate upon 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.

scholarly journals Engineering Zinc Finger Proteins using Accessory Binding Modules

2012 ◽  
Vol 102 (3) ◽  
pp. 74a
Author(s):  
Kathryn Trenshaw ◽  
Younghoon Kim ◽  
Nathan Yee ◽  
Peiyi Wang ◽  
Charles Schroeder
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Proteins

scholarly journals Identification of non-Ser/Thr-Pro consensus motifs for Cdk1 and their roles in mitotic regulation of C2H2 zinc finger proteins and Ect2

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Kazuhiro Suzuki ◽  
Kosuke Sako ◽  
Kazuhiro Akiyama ◽  
Michitaka Isoda ◽  
Chiharu Senoo ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Proteins ◽  
C2h2 Zinc Finger ◽  
Consensus Motifs ◽  
Mitotic Regulation

Negative regulation of the RTBV promoter by designed zinc finger proteins

2010 ◽  
Vol 72 (6) ◽  
pp. 621-630 ◽  
Author(s):  
M. Isabel Ordiz ◽  
Laurent Magnenat ◽  
Carlos F. Barbas ◽  
Roger N. Beachy
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Proteins ◽  
Negative Regulation

Drug discovery with engineered zinc-finger proteins

2003 ◽  
Vol 2 (5) ◽  
pp. 361-368 ◽  
Author(s):  
Andrew C. Jamieson ◽  
Jeffrey C. Miller ◽  
Carl O. Pabo
Keyword(s):  
Drug Discovery ◽  
Zinc Finger ◽  
Zinc Finger Proteins
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