CRISPR-Cpf1 Antibody (3D3-F7) [DyLight 550]

• Reactivity: Ba
• Applications: WB
• Clone: 3D3-F7
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: DyLight 550

CRISPR-Cpf1 Antibody (3D3-F7) [DyLight 550] Summary

• Immunogen: Partial recombinant protein made to a C-terminal portion of the Acidaminococcus sp. CRISPR-Cpf1 protein
• Specificity: This antibody is specific for the Acidaminococcus sp. CRISPR-Cpf1
• Isotype: IgG2a Kappa
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 50mM Sodium Borate
• Preservative: 0.05% Sodium Azide
• Purity: Protein A or G purified

Notes

DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

Alternate Names for CRISPR-Cpf1 Antibody (3D3-F7) [DyLight 550]

• Cas12a
• Clustered regularly interspaced short palindromic repeat (CRISPR) associated Cas12a
• Cpf1
• CRISPR-associated endonuclease Cas12a
• CRISPR-associated endonuclease Cpf1
• CRISPR-Cas12a
• FnCas12a
• FnCpf1

Background

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

Limitations

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.