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Application of CRISPR/Cas9 for biomedical discoveries.

Riordan SM, Heruth DP, Zhang LQ, Ye SQ - Cell Biosci (2015)

Bottom Line: The CRISPR/Cas9 system is much easier to customize and optimize because the site selection for DNA cleavage is guided by a short sequence of RNA rather than an engineered protein as in the systems of zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), and meganucleases.Although it still suffers from some off-target effects, the CRISPR/Cas9 system has been broadly and successfully applied for biomedical discoveries in a number of areas.We then present concise concluding remarks and future directions for this fast moving field.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, The Children's Mercy Hospital, Kansas City, MO USA.

ABSTRACT
The Clustered Regions of Interspersed Palindromic Repeats-Cas9 (CRISPR/Cas9), a viral defense system found in bacteria and archaea, has emerged as a tour de force genome editing tool. The CRISPR/Cas9 system is much easier to customize and optimize because the site selection for DNA cleavage is guided by a short sequence of RNA rather than an engineered protein as in the systems of zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), and meganucleases. Although it still suffers from some off-target effects, the CRISPR/Cas9 system has been broadly and successfully applied for biomedical discoveries in a number of areas. In this review, we present a brief history and development of the CRISPR system and focus on the application of this genome editing technology for biomedical discoveries. We then present concise concluding remarks and future directions for this fast moving field.

No MeSH data available.


History of CRISPR/Cas development. A useful understanding of the potential and capabilities of the CRISPR/Cas system took many years to develop. Only recently has there been an explosion in the development and application of this system
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Fig1: History of CRISPR/Cas development. A useful understanding of the potential and capabilities of the CRISPR/Cas system took many years to develop. Only recently has there been an explosion in the development and application of this system

Mentions: Although there is great diversity in the CRISPR/Cas systems, they all share three common stages: adaptation, crRNA biogenesis and immunity. Adaptation is the process of acquiring unique spacer sequences from viral and plasmid DNA. crRNA biogenesis involves the transcription and processing of crRNA. Immunity, also termed interference, entails the formation of an endonuclease complex capable of recognizing and digesting invading phage and plasmid DNA. The discovery of endonucleases that could be guided to specific regions of the genome was met with immediate interest. Successful attempts to develop easy to use genome editing methods lead to the creation of chimeric RNA systems containing crRNA and the trans activating RNA (tracrRNA) to create a single guide RNA (sgRNA) to be used with the Cas protein, Cas9 [23, 24]. Since then researchers at the Broad Institute of MIT and Harvard have been awarded a U.S. Patent for the technology, which they have licensed to multiple biotechnology companies leading to a huge growth in the technology and its availability. A summary of the history of CRISPR/Cas technological development can be seen in Fig. 1.Fig. 1


Application of CRISPR/Cas9 for biomedical discoveries.

Riordan SM, Heruth DP, Zhang LQ, Ye SQ - Cell Biosci (2015)

History of CRISPR/Cas development. A useful understanding of the potential and capabilities of the CRISPR/Cas system took many years to develop. Only recently has there been an explosion in the development and application of this system
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4487574&req=5

Fig1: History of CRISPR/Cas development. A useful understanding of the potential and capabilities of the CRISPR/Cas system took many years to develop. Only recently has there been an explosion in the development and application of this system
Mentions: Although there is great diversity in the CRISPR/Cas systems, they all share three common stages: adaptation, crRNA biogenesis and immunity. Adaptation is the process of acquiring unique spacer sequences from viral and plasmid DNA. crRNA biogenesis involves the transcription and processing of crRNA. Immunity, also termed interference, entails the formation of an endonuclease complex capable of recognizing and digesting invading phage and plasmid DNA. The discovery of endonucleases that could be guided to specific regions of the genome was met with immediate interest. Successful attempts to develop easy to use genome editing methods lead to the creation of chimeric RNA systems containing crRNA and the trans activating RNA (tracrRNA) to create a single guide RNA (sgRNA) to be used with the Cas protein, Cas9 [23, 24]. Since then researchers at the Broad Institute of MIT and Harvard have been awarded a U.S. Patent for the technology, which they have licensed to multiple biotechnology companies leading to a huge growth in the technology and its availability. A summary of the history of CRISPR/Cas technological development can be seen in Fig. 1.Fig. 1

Bottom Line: The CRISPR/Cas9 system is much easier to customize and optimize because the site selection for DNA cleavage is guided by a short sequence of RNA rather than an engineered protein as in the systems of zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), and meganucleases.Although it still suffers from some off-target effects, the CRISPR/Cas9 system has been broadly and successfully applied for biomedical discoveries in a number of areas.We then present concise concluding remarks and future directions for this fast moving field.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, The Children's Mercy Hospital, Kansas City, MO USA.

ABSTRACT
The Clustered Regions of Interspersed Palindromic Repeats-Cas9 (CRISPR/Cas9), a viral defense system found in bacteria and archaea, has emerged as a tour de force genome editing tool. The CRISPR/Cas9 system is much easier to customize and optimize because the site selection for DNA cleavage is guided by a short sequence of RNA rather than an engineered protein as in the systems of zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), and meganucleases. Although it still suffers from some off-target effects, the CRISPR/Cas9 system has been broadly and successfully applied for biomedical discoveries in a number of areas. In this review, we present a brief history and development of the CRISPR system and focus on the application of this genome editing technology for biomedical discoveries. We then present concise concluding remarks and future directions for this fast moving field.

No MeSH data available.