Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy

a.
State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, The Chinese Academy of Sciences, Datun Road 15, Beijing 100101, China
b.
Key Laboratory of Cell Proliferation and Differentiation of Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China
c.
Department of Biological Science, Florida State University, Tallahassee, Florida 32304-4295, USA

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Corresponding author: E-mail address: wumin@bio.fsu.edu (Wu-Min Deng); E-mail address: rjiao@sun5.ibp.ac.cn (Renjie Jiao)

摘要

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Abstract: Technology development has always been one of the forces driving breakthroughs in biomedical research. Since the time of Thomas Morgan, Drosophilists have, step by step, developed powerful genetic tools for manipulating and functionally dissecting the Drosophila genome, but room for improving these technologies and developing new techniques is still large, especially today as biologists start to study systematically the functional genomics of different model organisms, including humans, in a high-throughput manner. Here, we report, for the first time inDrosophila, a rapid, easy, and highly specific method for modifying the Drosophila genome at a very high efficiency by means of an improved transcription activator-like effector nuclease (TALEN) strategy. We took advantage of the very recently developed “unit assembly” strategy to assemble two pairs of specific TALENs designed to modify the yellow gene (on the sex chromosome) and a novel autosomal gene. The mRNAs of TALENs were subsequently injected into Drosophila embryos. From 31.2% of the injected F₀ fertile flies, we detected inheritable modification involving the yellow gene. The entire process from construction of specific TALENs to detection of inheritable modifications can be accomplished within one month. The potential applications of this TALEN-mediated genome modification method in Drosophila are discussed.
- TALEN /
- Genomic modification /
- Unit assembly /
- Reverse genetics

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doi: 10.1016/j.jgg.2012.04.003

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Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy

Corresponding author: E-mail address: wumin@bio.fsu.edu (Wu-Min Deng); E-mail address: rjiao@sun5.ibp.ac.cn (Renjie Jiao)

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doi: 10.1016/j.jgg.2012.04.003

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Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy

Corresponding author: E-mail address: wumin@bio.fsu.edu (Wu-Min Deng); E-mail address: rjiao@sun5.ibp.ac.cn (Renjie Jiao)

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