Comparison

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

Z03386-10

GenCrispr Cas9 Nuclease

10 μg (0.2mg/ml)

Z03386-50

GenCrispr Cas9 Nuclease

50 μg (5×10μg)(0.2mg/ml)

Z03385-50

GenCrispr Cas9-C-Nuclease

50 μg (1mg/ml)

Z03385-100

GenCrispr Cas9-C-Nuclease

100 μg (4mg/ml)

Z03388-50

GenCrispr Cas9-N-NLS Nuclease

50 µg (1mg/ml)

Z03388-100

GenCrispr Cas9-N-NLS Nuclease

100 µg (4mg/ml)

Z03389-50

GenCrispr NLS-Cas9-NLS Nuclease

50 µg (1mg/ml)

Z03389-100

GenCrispr NLS-Cas9-NLS Nuclease

100 µg (4mg/ml)

Z03390-100

GenCrispr NLS-Cas9-D10A Nickase

100 µg (4mg/ml)

Z03390-10

GenCrispr NLS-Cas9-D10A Nickase

10 µg (1mg/ml)

Z03390-50

GenCrispr NLS-Cas9-D10A Nickase

50 µg (1mg/ml)

Z03393-50

GenCrispr NLS-Cas9-EGFP Nuclease

50 µg (1mg/ml)

Z03393-100

GenCrispr NLS-Cas9-EGFP Nuclease

100 µg (3mg/ml)

 

Kits

Cat. No.

Product Name

Size

L00688-25

GenCrispr Mutation Detection Kit

25 reactions

L00688-100

GenCrispr Mutation Detection Kit

100 reactions

L00689-30

GenCrispr sgRNA Screening Kit

30 reactions

L00689-100

GenCrispr sgRNA Screening Kit

100 reactions

L00690-25

High-Efficiency sgRNA-Cas9-GFP Plasmid (linear) Assembly Kit

25 reactions

L00690-10

High-Efficiency sgRNA-Cas9-GFP Plasmid (linear) Assembly Kit

10 reactions

L00691-25

High-Efficiency gRNA-Cas9-Puro Plasmid (linear) Assembly Kit

25 reactions

L00691-10

High-Efficiency gRNA-Cas9-Puro Plasmid (linear) Assembly Kit

10 reactions

L00692-25

High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit

25 reactions

L00692-10

High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit

10 reactions

L00693-25

High-Efficiency gRNA-Cas9-GFP Plasmid (linear) Assembly Kit

25 reactions

L00693-10

High-Efficiency gRNA-Cas9-Puro Plasmid Assembly Kit

10 reactions

L00694-50

 High-Efficiency gRNA-Cas9-Puro Plasmid Assembly Kit

50 reactions

L00694-20

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 Immunology32: 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 biotechnology31(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 biotechnology34(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