GenScript’s offerings are cost-effective with reliable and proven performance for removal of contaminating nucleic acids. Furthermore, the Benz-Neburase’s offerings are produced under stringent quality controls with materials and manufacturing process that are fully traceable, allowing for support of IND filing and progression of development stages for various applications.
Recombinant
Cas12a with an C-terminal NLS expressed by E.coli
浓度
4 mg/ml
描述
The clustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonly present in archaea and bacteria. The CRISPR systems enable the host to specifically target and cleave foreign nucleic acids thus targeting infectiousviruses and plasmids. Recently, a type V CRISPR system has been identified in several bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an additional trans-activating crRNA (tracrRNA), and allow for targeting of additional genomic regions by cleaveing the target DNA proceeded by a short T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and purified. The nuclease contains nuclear localization sequence (NLS) at the C-terminus and 6× His-tag at the C-terminus.
GenCRISPR™ ErCas12a Nuclease is an RNA-guided DNA endonuclease from Eubacterium rectale. It recognizes a T-rich protospacer adjacent motif (PAM) and results in a staggered DNA double-strand break (DSB). After the specific cleavage, Cas12a can also activate collateral cleavage activity towards adjacent non-specific ssDNA sequences. Hence, Cas12a nuclease is a good alternative for Cas9 in certain target DNA editing, and provides a novel strategy for DNA detection.
GenCRISPR™ LbCas12a Nuclease is an RNA-guided DNA endonuclease from Lachnospiraceae bacterium. Cas12-family enzymes exhibit two DNase activity modes; besides crRNA-guided cis-cleavage of cDNA targets, these nucleases also catalyze trans-cleavage of non-target ssDNA substrates introduced by target DNA, which has been exploited to develop sensitive technologies for nucleic acid detection.
GenCRISPR Cas13a (C2c2) Nuclease is an RNA-guided, RNA-targeting CRISPR enzyme, commonly referred to as C2c2. This enzyme is a member of Type VI CRISPR family containing a single protein effector with two HEPN domains. The Cas13a exhibits two RNase activities. The first activity, a cis- sequence-specific RNA guided cleavage, activates the secondary RNase activity of the enzyme, a non-specific trans- ribonuclease activity. This Cas13a protein can be applied to detect single RNA molecules with proven high specificity.
GenCrispr NLS-Cas9-EGFP is a fusion protein developed by GenScript. It contains a nuclear localization signal (NLS) on its N terminal end, and an EGFP and a 6x(His) sequence on the C terminal end. Cas9 nuclease is an RNA-guided endonuclease that can catalyze cleavage of double stranded DNA. This kind of targeted nuclease is a powerful tool for genome editing with high precision. Cas9 protein forms a highly stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9 system. When Cas9 is expressed with an NLS sequence, the Cas9 RNP complex can localize to the nucleus immediately upon entering the cell. There is no requirement for in vivo transcription or translation, which improves the efficiency of this method over plasmid-based systems. Additionally, the Cas9 RNP complex is rapidly cleared from the cells minimizing the chance of off-target cleavage when compared to other systems (Kim, et al. 2014). The EGFP tag can be used as a reporter for tracking or sorting transfected cells, which enables enrichment of cell populations for desired genome edits via fluorescence activated cell sorting (FACS). It significantly reduces the labor and cost associated with single cell cloning and genotyping in genome editing applications.