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Zinc Finger Assembly

Rapid Assembly and Cloning of Zinc Finger Proteins with ...

Fluorescent Protein-Based Molecular Beacons by Zinc Finger Protein-Guided Assembly Daniel Blackstock, Qing Sun, Wilfred Chen Department of Chemical and Biomolecular Engineering, University of Delaware, Newark,

Highly active zinc-finger nucleases by extended modular ...

The modular assembly (MA) method of generating engineered zinc finger proteins (ZFPs) was the first practical method for creating custom DNA-binding proteins.

Zinc Finger Nuclease and Homing Endonuclease-Mediated ...

Joung and others have shown that, in practice, modular assembly has a very low success rate for creating multi-finger proteins. This high failure rate is most likely due to "context-dependent" effects that individual zinc fingers can have on the DNA-binding activities of their neighboring fingers.

Zinc finger nuclease - Wikipedia

assembly. A novel zinc-finger within the conserved domain, CX10CX11CX2H, is essential for the homocomplex formation. Mutant alleles with amino acid substitutions at conserved cysteines and histidine in the zinc-finger fail to assemble homocomplexes. Defect in homocomplex assembly …

ZiFiT

A zinc finger is a small protein structural motif that is characterized by the coordination of one or more zinc ions (Zn 2+) in order to stabilize the fold.Originally coined to describe the finger-like appearance of a hypothesized structure from Xenopus laevis transcription factor IIIA, the zinc finger name has now come to encompass a wide variety of differing protein structures.

Programed Assembly of Nucleoprotein Nanoparticles Using ...

Zinc Finger Tools offers a scoring function to assist users in choosing among potential zinc finger target sites, but as noted on the Zinc Finger Tools website itself: rigorous empirical validation of the scoring function awaits further experimental data.

The Zinc Finger Consortium | Protocols

OPEN zinc finger design selects zinc-fingers by screening an archive of characterized zinc fingers for each DNA triplet, followed by random PCR assembly and selection of the best assembled zinc finger …

A Novel Zinc-Finger Is Required for Mcm10 Homocomplex …

Addgene plasmid kits for cloning, genome engineering, fluorescent proteins & more. ... Zinc Finger Consortium: Modular Assembly Accessory Reagents Kit - Joung; Zinc Finger Consortium: OPEN Reagents - Joung ... Learn about the latest plasmid technologies …

The Generation of Zinc Finger Proteins by Modular Assembly

The CoDA method described by the Zinc Finger Consortium bypasses this problem by using two separate archives of two-finger modules that share common N-terminal or C-terminal fingers, which permits the assembly of three-finger proteins through overlap at these common units ( 52).

Center for Computational and Integrative Biology

Zinc-Finger Nucleases. The class of targeting reagents that has proved the most versatile and effective in recent years is that of the zinc-finger nucleases (ZFNs), which have separate DNA-binding and DNA-cleavage domains (Figures 3 and 4).

Zinc Finger Consortium Expression Vector and Modular ...

In the latter approach, the 5 positions at the finger-finger interaction interface of the selected modules were used as a single unit for zinc finger protein assembly, where the module for assembly spans from position 3 of the N-terminal finger to position 3 of the C-terminal finger within the two-finger module.

Addgene: Plasmid Kits

Zinc Finger Consortium Expression Vector and Modular Assembly Kit Description: The Zinc Finger Consortium () was established to promote continued research and development of engineered zinc finger technology. Consortium members Keith Joung and Daniel Voytas have deposited their Expression Vector Kit and Modular Assembly Kit ...

The Zinc Finger Consortium | Reagents

Kit #1000000005 includes 141 zinc finger plasmids encoding different zinc finger modules from the Sangamo, Barbas, and Toolgen module sets, and can be used to efficiently assemble zinc finger arrays via modular assembly. Also included in this kit are 5 plasmids and a strain for the bacterial two-hybrid (B2H) system used to screen zinc finger ...

Zinc Finger Nucleases as tools to understand and treat ...

Scheme 1. Rapid assembly of zinc finger protein based on the Gibson assembly. PCR primers to amplify one-finger module have a variable region at the 5' end and a common region at the 3' end. The variable region encodes an α-helix region of the zinc finger, which is responsible for specific interaction with DNA bases.

Genome Engineering With Zinc-Finger Nucleases | Genetics

Context-Dependent Assembly (CoDA) method for engineering zinc finger arrays Oligomerized Pool Engineering (OPEN) method for engineering zinc finger arrays ; Modular Assembly method for engineering zinc finger arrays

Clinical Scale Zinc Finger Nuclease-mediated Gene Editing ...

The modular assembly (MA) method of generating engineered zinc finger proteins (ZFPs) was the first practical method for creating custom DNA-binding proteins. As such, MA has enabled a vast exploration of sequence-specific methods and reagents, ushering in the modern era of zinc finger-based applications that are described in this volume.

Evaluation and application of modularly assembled zinc ...

Engineered zinc finger nucleases (ZFNs) are a tool for genome manipulation that are of great interest to scientists in many fields. To meet the needs of researchers wishing to employ ZFNs, an inexpensive, rapid assembly procedure would be beneficial to laboratories that do not have access to the ...

C. elegans PAT-9 is a nuclear zinc finger protein critical ...

ZiFiT also continues to support Zinc Finger Engineering (Versions 3.2 & 3.3) for the CoDA (Context-Dependent Assembly; Sander et al., Nat. Methods 2011) and OPEN (Oligomerized Pool Engineering; Maeder et al., Mol Cell 2008) design methods developed by Keith Joung's lab at the Massachusetts General Hospital and Harvard Medical School.

Synthetic Zinc Finger Nuclease Design and Rapid Assembly ...

Nov 19, 2018· Zinc finger domains are found in some viral proteins, including the neocapsid (NC) protein of the Human Immunodeficiency-1 (HIV-1) virus. NC is an attractive antiviral target because it is important for viral assembly and is highly conserved.

The generation of zinc finger proteins by modular assembly

The creation of zinc finger motif-fused enzymes thus offers a promising option for the assembly of enzymes, with a resulting proximity effect, onto a DNA scaffold directly without the creation of ssDNA–enzyme conjugate beforehand.

Zinc Finger Targeter (ZiFiT): an engineered zinc finger ...

Zinc-finger nucleases (ZFNs) are important tools for genome engineering. Despite intense interest by many academic groups, the lack of robust noncommercial methods has hindered their widespread use. The modular assembly (MA) of ZFNs from publicly available one-finger …

Using defined finger–finger interfaces as units of ...

Zinc Finger Consortium Expression Vector and Modular Assembly Kit Description: The Zinc Finger Consortium () was established to promote continued research and development of engineered zinc finger technology. Consortium members Keith Joung and Daniel Voytas have deposited their Expression Vector Kit and Modular Assembly Kit ...

The Generation of Zinc Finger Proteins by Modular Assembly ...

The modular assembly (MA) method of generating engineered zinc finger proteins (ZFPs) was the first practical method for creating custom DNA-binding proteins. As such, MA has enabled a vast exploration of sequence-specific methods and reagents, ushering in the modern era of zinc finger-based ...

ZiFiT

Nov 21, 2008· Broad applications of zinc finger nuclease (ZFN) technology—which allows targeted genome editing—in research, medicine, and biotechnology are hampered by the lack of a convenient, rapid, and publicly available method for the synthesis of functional ZFNs.

Addgene: Zinc Finger Consortium - Modular Assembly Kits

Schematic representation of the modified tumor infiltrating lymphocytes (TIL) process and zinc finger nuclease (ZFN) target site at the PD-1 locus. (a) Schematic of the ZFN target site within the PD-1 gene at the 2q37.3 location. Each ZFN comprises five zinc finger domains that recognize a composite 15bp target site (in red).

Modular Zinc Finger Assembly protocol revised

Thirty-six three-finger arrays (named "ZFA __" for zinc finger array) were assembled using modules based on the Zinc Finger Consortium Modular Assembly Kit 1.0,1 and the identities of the three modules used to construct each three-finger array use the Consortium

Zinc Finger Consortium Expression Vector and Modular ...

This site includes information about the Zinc Finger Consortium, including: zinc fingers, Cys2His2 zinc finger, C2H2 zinc finger, C2H2 ZF, ZF, ZFN, ZFC, zinc finger ...

Targeted genome editing in human cells with zinc finger ...

Modular Assembly to generate site-specific zinc finger nucleases (ZFN) This protocol entails two separate PCR steps: first, to separately amplify each of the 3 individual

Zinc Finger Nuclease and Homing Endonuclease-Mediated ...

Other procedures can utilize either 1-finger or 2-finger modules to generate zinc-finger arrays with six or more individual zinc fingers. The main drawback with this procedure is the specificities of individual zinc fingers can overlap and can depend on the context of the surrounding zinc fingers and DNA.

Assembly of zinc finger motif-fused enzymes on a dsDNA ...

Zinc finger nuclease (ZFN) is a chimeric restriction enzyme made of a C 2 H 2-type zinc finger protein (ZFP) and the FokI nuclease domain (F N).ZFN technology has been considered as a …