Site-directed transposon integration in human cells.

Site-directed transposon integration in human cells. Research Abstract Details 

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Site-directed transposon integration in human cells. Abstract Text:

Stephen R Yant,Yong Huang,Bassel Akache,Mark A Kay,

The Sleeping Beauty (SB) transposon is a promising gene transfer vector that integrates nonspecifically into host cell genomes. Herein, we attempt to direct transposon integration into predetermined DNA sites by coupling a site-specific DNA-binding domain (DBD) to the SB transposase. We engineered fusion proteins comprised of a hyperactive SB transposase (HSB5) joined via a variable-length linker to either end of the polydactyl zinc-finger protein E2C, which binds a unique sequence on human chromosome 17. Although DBD linkage to the C-terminus of SB abolished activity in a human cell transposition assay, the N-terminal addition of the E2C or Gal4 DBD did not. Molecular analyses indicated that these DBD-SB fusion proteins retained DNA-binding specificity for their respective substrate molecules and were capable of mediating bona fide transposition reactions. We also characterized transposon integrations in the presence of the E2C-SB fusion protein to determine its potential to target predefined DNA sites. Our results indicate that fusion protein-mediated tethering can effectively redirect transposon insertion site selection in human cells, but suggest that stable docking of integration complexes may also partially interfere with the cut-and-paste mechanism. These findings illustrate the feasibility of directed transposon integration and highlight potential means for future development.

Site-directed transposon integration in human cells. Publishing Authors By Initials

SR Yant,Y Huang,B Akache,MA Kay,

For similar enzymes and coenzymes: enzymes: recombinases: integrases: transposases research abstracts see: enzymes and coenzymes: enzymes: recombinases: integrases: transposases research

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Site-directed transposon integration in human cells. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Nucleic acids research

VOLUME: 35

Page Numbers: e50

Journal Abbreviation:

ISSN: 1362-4962

DAY: 7

MONTH: 03

YEAR: 2007

Site-directed transposon integration in human cells. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 411011

Site-directed transposon integration in human cells. Keywords Mesh Terms:

KEYWORDS: Transposases

MESH TERMS: metabolism

Chemical & Substance for Abstract: Site-directed transposon integration in human cells. Information

Substance Name: sleeping beauty transposase, human

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Site-directed transposon integration in human cells.

AFFILIATION: Department of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305-5208, USA.

Country: England

AGENCY: United States NIDDK

GRANT: DK49022

ACRONYM: DK

MEDLINETA: Nucleic Acids Res

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