Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 Jan 24;1(1):e3.
doi: 10.1038/mtna.2011.5.

Targeting DNA With Fingers and TALENs

Daniel F Carlson  1 , Scott C Fahrenkrug, Perry B Hackett
Affiliations

Targeting DNA With Fingers and TALENs

Daniel F Carlson et al. Mol Ther Nucleic Acids. .
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
Comparison of zinc-finger nuclease (ZFN) and Transcription Activator-Like Effector Nuclease (TALEN) architecture. (a) ZFNs. Each ZFN polypeptide consists of two functional domains, a DNA-binding domain comprising a chain of finger modules (ZFs) that each typically recognize a unique 3-base pair sequence of DNA and a DNA-cleaving domain composed of the nuclease domain of the FokI nuclease. FokI functions as a dimer, hence when two FokI nucleases bind to DNA proximal to one another they can dimerize and introduce a double-strand break. Targeted double-strand DNA cutting can be obtained by designing zinc fingers for specific sequences that flank the desired cleavage site; in the example 12 base pairs per ZFN are targeted with polypeptides containing four zinc-finger modules each (ZF-1 through ZF-4 and ZF-5 through ZF-8). (b) Model of a TALEN. A TAL Effector (TALE) polypeptide contains a series of typically 34-amino acid repeats, of which residues 12 and 13 [repeat variable diresidues (RVDs) shown in orange] are responsible for recognition of a specific base as shown in the box (note that there is some discussion about the precision of the RVD NK recognition of G and other RVDs can specify base contacts). FokI nuclease is fused to the C-terminal end of the protein using wild-type TALE sequence as a spacer. Several spacer lengths between the TALE binding core and FokI have demonstrated activity. The number of tandem 34-amino acid repeats in the binding core defines the length of the recognition sequence, and the end of the functional DNA-binding motif. Each target sequence must be preceded by a T nucleotide. Two TALENs are shown to assemble on a genomic sequence in the opposite polarity to ZFNs to form a heterodimeric cleavage complex.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Rouet P, Smih F., and, Jasin M. Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells. Proc Natl Acad Sci USA. 1994;91:6064–6068. - PMC - PubMed
    1. Smih F, Rouet P, Romanienko PJ., and, Jasin M. Double-strand breaks at the target locus stimulate gene targeting in embryonic stem cells. Nucleic Acids Res. 1995;23:5012–5019. - PMC - PubMed
    1. Kim YG, Cha J., and, Chandrasegaran S. Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain. Proc Natl Acad Sci USA. 1996;93:1156–1160. - PMC - PubMed
    1. Porteus MH., and, Baltimore D. Chimeric nucleases stimulate gene targeting in human cells. Science. 2003;300:763. - PubMed
    1. Bibikova M, Beumer K, Trautman JK., and, Carroll D. Enhancing gene targeting with designed zinc finger nucleases. Science. 2003;300:764. - PubMed

LinkOut - more resources