Cas9 and Associated Proteins Antibody Sampler Kit #73211
Product Information
Kit Usage Information
Protocols
- 7074: Western Blotting
- 19984: Western Blotting, Immunofluorescence, Flow
- 51610: Western Blotting, Immunofluorescence, Flow
- 65832: Western Blotting, IF-F Citrate Retrieval (Rabbit), Immunofluorescence*, Flow
- 90111: Western Blotting, Immunofluorescence, Flow
Product Description
Specificity / Sensitivity
Each antibody in the Cas9 and Associated Proteins Antibody Sampler Kit recognizes transfected levels of its target protein. Antibodies are specific for the indicated endonuclease target and cross-reactivity with other endonucleases is not observed.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Val16 of Cas9 (S. pyogenes), Val905 of Cas9 (S. aureus), Leu822 of Acidaminococcus sp. Cpf1 (Strain BV3L6), and Ile841 of Cpf1 from Francisella tularensis subsp. novicida (Strain U112).
Background
CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are RNA-guided nuclease effectors that are utilized for precise genome editing in mammalian systems (1). Class 2 CRISPR systems rely on single-component effector proteins to mediate DNA interference (2). Several Class 2 CRISPR effector proteins, derived from specific bacterial species, are used for genome editing. Cas9 family of proteins, derived from S. pyogenes and S. aureus, are some of the most well characterized and widely used editing effector enzymes. Additional members of the Class2 CRISPR system include Cpf1/Cas12a (CRISPR from Prevotella and Francisella) endonucleases (3). Cpf1/Cas12a endonucleases, compared to Cas9 systems, have several unique features that increase the utility of CRISPR-based genome editing techniques: 1) Cpf1/Cas12a-mediated cleavage relies on a single and short CRISPR RNA (crRNA) without the requirement of a trans-activating crRNA (tracrRNA), 2) Cpf1/Cas12a utilizes T-Rich protospacer adjacent motif (PAM) sequences rather than a G-Rich PAM, and 3) Cpf1/Cas12a generates a staggered, rather than a blunt-ended, DNA double-stranded break (3). These features broaden the utility of using CRISPR-Cas systems for specific gene regulation and therapeutic applications. Several Cpf1/Cas12a bacterial orthologs, e.g. Francisella novicida U112 and Acidaminococcus sp. BV3L6, have been characterized for CRISPR-mediated mammalian genome editing (3,4).
Limited Uses
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