TY - JOUR
T1 - Comparing Zinc Finger Nucleases and Transcription Activator-Like Effector Nucleases for Gene Targeting in Drosophila
AU - Beumer, Kelly J.
AU - Trautman, Jonathan K.
AU - Christian, Michelle
AU - Dahlem, Timothy J.
AU - Lake, Cathleen M.
AU - Hawley, R. Scott
AU - Grunwald, David J.
AU - Voytas, Daniel F.
AU - Carroll, Dana
PY - 2013
Y1 - 2013
N2 - Zinc-nger nucleases have proven to be successful as reagents for targeted genome manipulation in Drosophila melanogaster and many other organisms. Their utility has been limited, how-ever, by the signicant failure rate of new designs, reecting the complexity of DNA recognition by zinc ngers. Transcription activator-like effector (TALE) DNA-binding domains depend on a simple, one-module-to-one-base-pair recognition code, and they have been very productively incorporated into nucleases (TALENs) for genome engineering. In this report we describe the design of TALENs for a number of different genes in Drosophila, and we explore several parameters of TALEN design. The rate of success with TALENs was substantially greater than for zinc-nger nucleases, and the frequency of mutagenesis was comparable. Knockout mutations were isolated in several genes in which such alleles were not previously available. TALENs are an effective tool for targeted genome manipulation in Drosophila.
AB - Zinc-nger nucleases have proven to be successful as reagents for targeted genome manipulation in Drosophila melanogaster and many other organisms. Their utility has been limited, how-ever, by the signicant failure rate of new designs, reecting the complexity of DNA recognition by zinc ngers. Transcription activator-like effector (TALE) DNA-binding domains depend on a simple, one-module-to-one-base-pair recognition code, and they have been very productively incorporated into nucleases (TALENs) for genome engineering. In this report we describe the design of TALENs for a number of different genes in Drosophila, and we explore several parameters of TALEN design. The rate of success with TALENs was substantially greater than for zinc-nger nucleases, and the frequency of mutagenesis was comparable. Knockout mutations were isolated in several genes in which such alleles were not previously available. TALENs are an effective tool for targeted genome manipulation in Drosophila.
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U2 - 10.1534/g3.113.007260
DO - 10.1534/g3.113.007260
M3 - Article
C2 - 23979928
AN - SCOPUS:84885791114
SN - 2160-1836
VL - 3
SP - 1717
EP - 1725
JO - G3: Genes, Genomes, Genetics
JF - G3: Genes, Genomes, Genetics
IS - 9
ER -