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Mouse Anti-CRISPR-Cas9 Recombinant Antibody (0053) (V2LY-1206-LY1174)


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Published Data
V2LY-1206-LY1174-1
Mouse Anti-CRISPR-Cas9 Recombinant Antibody (6G12) (V2LY-1206-LY1174) in WB
Western blot for SaCas9 protein expression. Protein expression for SaCas9 was shown in the positive control as well as in A17L and I2L targets 48 h post-infection. Protein expression was not detected for either targets at 72 h post-infection. ...View More
Siegrist, C. M., Kinahan, S. M., Settecerri, T., Greene, A. C., & Santarpia, J. L. (2020). CRISPR/Cas9 as an antiviral against Orthopoxviruses using an AAV vector. Scientific reports, 10(1), 19307.
Distributed under Open Access license CC BY 4.0, without modification.

Summary

Host Animal
Mouse
Specificity
Staphylococcus aureus
Clone
0053
Antibody Isotype
IgG2b, κ
Application
WB, IP, IF, ELISA

Basic Information

Immunogen
His-tagged recombinant Cas-9 from Staphylococcus aureus.
Host Species
Mouse
Specificity
Staphylococcus aureus
Antibody Isotype
IgG2b, κ
Clonality
Monoclonal Antibody
Application Notes
ApplicationNote
WB1:500-1:2,000
IP1-2 µg/10^6 cells
IF(ICC)1:500-1:2,000
ELISA1:1,000-1:2,000

Formulations & Storage [For reference only, actual COA shall prevail!]

Format
Liquid
Buffer
Tris-Glycine
Preservative
0.05% sodium azide
Concentration
Batch dependent
Purity
>95% as determined by analysis by SDS-PAGE
Storage
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freezethaw cycles.

Target

Full Name
CRISPR-associated endonuclease Cas9
Function
CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids) (PubMed:21455174).

CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA; Cas9 only stabilizes the pre-crRNA:tracrRNA interaction and has no catalytic function in RNA processing (PubMed:24270795).

Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed 3'-5' exonucleolytically. DNA-binding requires protein and both gRNAs, as does nuclease activity. Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. DNA strand separation and heteroduplex formation starts at PAM sites; PAM recognition is required for catalytic activity (PubMed:24476820).

Confers immunity against a plasmid with homology to the appropriate CRISPR spacer sequences (CRISPR interference) (PubMed:21455174).
Biological Process
Defense response to virus Source: UniProtKB-UniRule
Maintenance of CRISPR repeat elements Source: UniProtKB
More Infomation

Yip, B. H. (2020). Recent advances in CRISPR/Cas9 delivery strategies. Biomolecules, 10(6), 839.

Zhan, T., Rindtorff, N., Betge, J., Ebert, M. P., & Boutros, M. (2019, April). CRISPR/Cas9 for cancer research and therapy. In Seminars in cancer biology (Vol. 55, pp. 106-119). Academic Press.

You, L., Tong, R., Li, M., Liu, Y., Xue, J., & Lu, Y. (2019). Advancements and obstacles of CRISPR-Cas9 technology in translational research. Molecular Therapy-Methods & Clinical Development, 13, 359-370.

Cui, Y., Xu, J., Cheng, M., Liao, X., & Peng, S. (2018). Review of CRISPR/Cas9 sgRNA design tools. Interdisciplinary Sciences: Computational Life Sciences, 10(2), 455-465.

Harrington, L. B., Doxzen, K. W., Ma, E., Liu, J. J., Knott, G. J., Edraki, A., ... & Doudna, J. A. (2017). A broad-spectrum inhibitor of CRISPR-Cas9. Cell, 170(6), 1224-1233.

Jiang, F., & Doudna, J. A. (2017). CRISPR–Cas9 structures and mechanisms. Annual review of biophysics, 46, 505-529.

Liu, X., Wu, S., Xu, J., Sui, C., & Wei, J. (2017). Application of CRISPR/Cas9 in plant biology. Acta pharmaceutica sinica B, 7(3), 292-302.

Pulecio, J., Verma, N., Mejía-Ramírez, E., Huangfu, D., & Raya, A. (2017). CRISPR/Cas9-based engineering of the epigenome. Cell Stem Cell, 21(4), 431-447.

Tang, L., Zeng, Y., Du, H., Gong, M., Peng, J., Zhang, B., ... & Liu, J. (2017). CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein. Molecular genetics and genomics, 292(3), 525-533.

Rauch, B. J., Silvis, M. R., Hultquist, J. F., Waters, C. S., McGregor, M. J., Krogan, N. J., & Bondy-Denomy, J. (2017). Inhibition of CRISPR-Cas9 with bacteriophage proteins. Cell, 168(1-2), 150-158.

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For research use only. Not intended for any clinical use.

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