CRISPR-Cpf1 Antibody [FITC]

Novus Biologicals | Catalog # NBP2-62735F

Novus Biologicals
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Key Product Details

Species Reactivity

Bacterial

Applications

Western Blot

Label

FITC (Excitation = 495 nm, Emission = 519 nm)

Antibody Source

Polyclonal Rabbit IgG
View all Cpf1 Primary Antibodies »

Product Specifications

Immunogen

Partial recombinant Francisella tularensis protein (between amino acids 600-850) [NCBI WP_003034647.1]

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Applications for CRISPR-Cpf1 Antibody [FITC]

Application
Recommended Usage

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: Cpf1

Clustered regularly interspaced short palindromic repeats (CRISPRs) are derived from DNA fragments of bacteriophages that infect prokaryotes. When infected, the bacteria capture snips of DNA from the invading virus generating CRISPR arrays. During subsequent infections, the bacteria produce RNA segments from the CRISPR arrays to target the virus' DNA. CRISPR-associated protein 9 (Cas9) from Streptococcus pyogenes is a class 2/type II RNA-guided, DNA binding, and DNA cleaving enzyme. Cas9 functions as an integral component of the bacterial CRISPR adaptive immune system which targets and disables the virus' DNA (1, 2). To check for sites complementary to the 20 base pair spacer region of the guide RNA (gRNA) within the CRISPR, Cas9 unwinds foreign DNA that invades the bacteria. If the foreign DNA sequence is complementary to the gRNA, Cas9 cleaves the invading DNA, rendering the virus disabled. The presence of a 5'-NGG-3' protospacer adjacent motif (PAM) immediately downstream of the target DNA is required for Cas9 cleavage of foreign DNA.

Cpf1 represents a new generation of CRISPR effector nucleases first isolated from Prevotella and Francisella1 (3). Cpf1 or Cas12a is present in both bacteria and archea and is a class 2/type V CRISPR-Cas effector protein which shares some homology with Cas9. For example, Cpf1 and Cas 9 share homologous RuvC-like nuclease domain and Arg-rich region (4). However, key differences between CRISPR/Cpf1 and CRISPR/Cas9 have been noted. Cpf1 requires a 5'-TTTN-3' PAM sequence located upstream to the target DNA and generates a staggered DNA overhang upon cleavage. Additionally, Cpf1 possesses RNase activity which simplifies crRNA or gRNA processing (2).

Using CRISPR-Cas technology, double-stranded DNA breaks may be induced within specific targeted genome sequences (target DNA; protospacer) for insertion or removal of DNA sequences for gene editing applications. To target a specific genomic sequence, gRNAs complementary to specific DNA targets are created. The gRNA will recognize the DNA sequence, and the Cas9 or Cpf1/Cas12a enzyme will cleave the DNA at the targeted location. Once the targeted DNA is removed, the cell's own DNA repair mechanism inserts or removes a DNA sequence resulting in genomic editing. Cpf1 orthologs Acidaminococcus sp Cpf1 (AsCpf1 151kDa) and Lachnospiraceae bacterium Cpf1 (LbCpf1 143kDa) are able to achieve efficient gene editing in human cells (3).

References

1. Oakes, B. L., Fellmann, C., Rishi, H., Taylor, K. L., Ren, S. M., Nadler, D. C.,... Savage, D. F. (2019). CRISPR-Cas9 Circular Permutants as Programmable Scaffolds for Genome Modification. Cell, 176(1-2), 254-267.e216. doi:10.1016/j.cell.2018.11.052

2. Verwaal, R., Buiting-Wiessenhaan, N., Dalhuijsen, S., & Roubos, J. A. (2018). CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae. Yeast. https://doi.org/10.1002/yea.3278

3. Safari, F., Zare, K., Negahdaripour, M., Barekati-Mowahed, M., & Ghasemi, Y. (2019). CRISPR Cpf1 proteins: Structure, function and implications for genome editing. Cell and Bioscience. https://doi.org/10.1186/s13578-019-0298-7

4. Makarova, K. S., Wolf, Y. I., Alkhnbashi, O. S., Costa, F., Shah, S. A., Saunders, S. J.,... Koonin, E. V. (2015). An updated evolutionary classification of CRISPR-Cas systems. Nature Reviews Microbiology. https://doi.org/10.1038/nrmicro3569

Long Name

CRISPR-associated Endonuclease Cas12a

Alternate Names

Cas12a

Additional Cpf1 Products

Product Documents for CRISPR-Cpf1 Antibody [FITC]

Certificate of Analysis

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Product Specific Notices for CRISPR-Cpf1 Antibody [FITC]

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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