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CRISPR/CAS 9 - Info
What is CRISPR/CAS 9?
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats. The CRISPR/Cas system can be found in the Genome of prokaryotes and was first identified in Escherichia coli (E. coli) [1] . Parts of the CRISPR/Cas System contain sequences of DNA from viruses, termed ‘spacers’, which have infected the bacterium in the past. These spacers are then transcribed and processed into short CRISPR RNA (crRNA). These crRNAs bind to trans-activating crRNAs (tracrRNAs) and promote cleavage and silencing of pathogenic DNA by the CRISPR-associated protein (Cas) which acts as an endonuclease. Therefore the CRISPR/Cas system is part of the adaptive immune system of the bacterium and provides acquired immunity against pathogens [2].
Figure 1. Meachnism of CRIPR-mediated immunity in bacteria
Applications
The target recognition and therefore the cleavage and slicing of target DNA by the endonuclease only occurs when there is a conserved dinucleotide-containing protospacer adjacent motif (PAM) sequence upstream of the crRNA-binding region. This way of target identification ensures a highly accurate targeting.
The described structure of the CRISPR/Cas System makes it possible to be retargeted. Therefore CRISPR/Cas can be used to cleave virtually any DNA sequence by redesigning the crRNA [3]. This makes CRISPR/Cas9 a versatile genome editing tool for genetic modification of the target host genome.
CRISPR/Cas9 is often superior to traditional genome editing applications like Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), because it provides an easy and efficient approach to manipulate the genome [3]. Its benefits are used in a broad spectrum of scientific fields, such as: medicine and biology, pharmacology and biotechnology engineering [4].
Figure 2. Illustration of Cas9 nuclease programmed by the crRNA:tracr complex cutting both strands of genomic DNA 5' of the PAM
http://dharmacon.gelifesciences.com/gene-editing/crispr-cas9/crispr-guide-rna/
Products
Enzymes
|
Cat. No. |
Product Name |
Size |
|
GenCrispr Cas9 Nuclease |
10 μg (0.2mg/ml) |
|
|
GenCrispr Cas9 Nuclease |
50 μg (5×10μg)(0.2mg/ml) |
|
|
GenCrispr Cas9-C-Nuclease |
50 μg (1mg/ml) |
|
|
GenCrispr Cas9-C-Nuclease |
100 μg (4mg/ml) |
|
|
GenCrispr Cas9-N-NLS Nuclease |
50 µg (1mg/ml) |
|
|
GenCrispr Cas9-N-NLS Nuclease |
100 µg (4mg/ml) |
|
|
GenCrispr NLS-Cas9-NLS Nuclease |
50 µg (1mg/ml) |
|
|
GenCrispr NLS-Cas9-NLS Nuclease |
100 µg (4mg/ml) |
|
|
GenCrispr NLS-Cas9-D10A Nickase |
100 µg (4mg/ml) |
|
|
GenCrispr NLS-Cas9-D10A Nickase |
10 µg (1mg/ml) |
|
|
GenCrispr NLS-Cas9-D10A Nickase |
50 µg (1mg/ml) |
|
|
GenCrispr NLS-Cas9-EGFP Nuclease |
50 µg (1mg/ml) |
|
|
GenCrispr NLS-Cas9-EGFP Nuclease |
100 µg (3mg/ml) |
Kits
|
Cat. No. |
Product Name |
Size |
|
GenCrispr Mutation Detection Kit |
25 reactions |
|
|
GenCrispr Mutation Detection Kit |
100 reactions |
|
|
GenCrispr sgRNA Screening Kit |
30 reactions |
|
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GenCrispr sgRNA Screening Kit |
100 reactions |
|
|
High-Efficiency sgRNA-Cas9-GFP Plasmid (linear) Assembly Kit |
25 reactions |
|
|
High-Efficiency sgRNA-Cas9-GFP Plasmid (linear) Assembly Kit |
10 reactions |
|
|
High-Efficiency gRNA-Cas9-Puro Plasmid (linear) Assembly Kit |
25 reactions |
|
|
High-Efficiency gRNA-Cas9-Puro Plasmid (linear) Assembly Kit |
10 reactions |
|
|
High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit |
25 reactions |
|
|
High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit |
10 reactions |
|
|
High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit |
25 reactions |
|
|
High-Efficiency gRNA-Cas9-Puro Plasmid Assembly Kit |
10 reactions |
|
|
High-Efficiency gRNA-Cas9-Puro Plasmid Assembly Kit |
50 reactions |
|
|
GenCrispr sgRNA Synthesis Kit |
20 reactions |
References
1. Barrangou R (2015). "The roles of CRISPR-Cas systems in adaptive immunity and beyond". Current Opinion in Immunology. 32: 36–41
http://www.sciencedirect.com/science/article/pii/S0952791514001563
2. Gaj, T., Gersbach, C. A., & Barbas, C. F. (2013). ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Trends in biotechnology, 31(7), 397-405.
http://www.sciencedirect.com/science/article/pii/S0167779913000875
3. Barrangou, R., & Doudna, J. A. (2016). Applications of CRISPR technologies in research and beyond. Nature biotechnology, 34(9), 933-941.
https://www.nature.com/nbt/journal/v34/n9/abs/nbt.3659.html
4. Crauciuc, Andrei, et al. "Development, Applications, Benefits, Challenges and Limitations of the New Genome Engineering Technique. An Update Study." Acta Medica Marisiensis 63.1 (2017): 4-9.
https://www.degruyter.com/view/j/amma.2017.63.issue-1/amma-2017-0007/amma-2017-0007.xm
