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Generation of knockout rabbits using transcription activator-like effector nucleases.

Wang Y, Fan N, Song J, Zhong J, Guo X, Tian W, Zhang Q, Cui F, Li L, Newsome PN, Frampton J, Esteban MA, Lai L - Cell Regen (Lond) (2014)

Bottom Line: Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research.They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci.This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Guangzhou, 510530 China.

ABSTRACT
Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research. They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases have been employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively. This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbits using transcription activator-like effector nucleases, and a perspective of the field.

No MeSH data available.


Related in: MedlinePlus

Photographs of holding and injection pipettes, microforge, pipette puller and micromanipulator. A. Holding pipette. B. Injection pipette. C. Microforge. D. Pipette puller. E. Micromanipulator. The holding and injection pipettes can be observed on both sides of the micromanipulator.
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Fig4: Photographs of holding and injection pipettes, microforge, pipette puller and micromanipulator. A. Holding pipette. B. Injection pipette. C. Microforge. D. Pipette puller. E. Micromanipulator. The holding and injection pipettes can be observed on both sides of the micromanipulator.

Mentions: Holding pipette (Figure 4A). Pull out the borosilicate glass capillaries on the flame of an alcohol lamp, then break at the tip leaving an outside diameter of ~120-180 μm. Blunt the broken end until the inner diameter is reduced to ~20-30 μm. Bend the pipette close to the blunt end (about 300 μm back) at 30 degrees with a microforge (Figure 4C).


Generation of knockout rabbits using transcription activator-like effector nucleases.

Wang Y, Fan N, Song J, Zhong J, Guo X, Tian W, Zhang Q, Cui F, Li L, Newsome PN, Frampton J, Esteban MA, Lai L - Cell Regen (Lond) (2014)

Photographs of holding and injection pipettes, microforge, pipette puller and micromanipulator. A. Holding pipette. B. Injection pipette. C. Microforge. D. Pipette puller. E. Micromanipulator. The holding and injection pipettes can be observed on both sides of the micromanipulator.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230510&req=5

Fig4: Photographs of holding and injection pipettes, microforge, pipette puller and micromanipulator. A. Holding pipette. B. Injection pipette. C. Microforge. D. Pipette puller. E. Micromanipulator. The holding and injection pipettes can be observed on both sides of the micromanipulator.
Mentions: Holding pipette (Figure 4A). Pull out the borosilicate glass capillaries on the flame of an alcohol lamp, then break at the tip leaving an outside diameter of ~120-180 μm. Blunt the broken end until the inner diameter is reduced to ~20-30 μm. Bend the pipette close to the blunt end (about 300 μm back) at 30 degrees with a microforge (Figure 4C).

Bottom Line: Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research.They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci.This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Guangzhou, 510530 China.

ABSTRACT
Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research. They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases have been employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively. This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbits using transcription activator-like effector nucleases, and a perspective of the field.

No MeSH data available.


Related in: MedlinePlus