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ArciTect™ Cas9-eGFP Nuclease

Enhanced green fluorescent protein (eGFP)-tagged Cas9 nuclease for the generation of double-strand breaks in CRISPR-Cas9 genome editing

ArciTect™ Cas9-eGFP Nuclease

Enhanced green fluorescent protein (eGFP)-tagged Cas9 nuclease for the generation of double-strand breaks in CRISPR-Cas9 genome editing

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Enhanced green fluorescent protein (eGFP)-tagged Cas9 nuclease for the generation of double-strand breaks in CRISPR-Cas9 genome editing
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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

ArciTect™ Cas9-eGFP Nuclease is a fusion protein consisting of enhanced green fluorescent protein (eGFP) and the wild-type Cas9 recombinant protein from Streptococcus pyogenes. ArciTect™ Cas9-eGFP Nuclease contains a C-terminal-linked eGFP molecule. ArciTect™ Cas9-eGFP Nuclease requires association with a guide RNA—e.g. ArciTect™ sgRNA (Catalog #200-0013) or a duplex composed of ArciTect™ tracrRNA (Catalog #76016) and ArciTect™ crRNA (Catalog #76010)—to form a ribonucleoprotein (RNP) complex. This RNP complex creates double-strand breaks at site-specific locations in the genome. ArciTect™ Cas9-eGFP Nuclease also contains a nuclear localization signal at the N-terminus, ensuring that the RNP complex translocates to the nucleus, thereby increasing the efficiency of genome editing. As the RNP complex is fully functional upon transfection, it allows for immediate activity following translocation to the nucleus. The RNP complex is degraded over 48 hours, allowing sufficient time for genome editing to occur while reducing off-target effects that can be caused by the continuous presence of the RNP complex. Using the RNP system also circumvents the laborious process of generating stable Cas9-expressing cell lines, saving time and reducing the risk of off-target effects due to leaky inducible expression systems. The S. pyogenes Cas9 uses the protospacer adjacent motif (PAM) sequence NGG (where N can be any nucleotide). The enzyme will not cleave without a genomic PAM site downstream of the target sequence
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Genome Editing
Brand
ArciTect
Area of Interest
Stem Cell Biology

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Name
Catalog #
76006
Lot #
All
Language
English
Document Type
Product Name
Catalog #
76006
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Publications (1)

Chamber-specific chromatin architecture guides functional interpretation of disease-associated Cis-regulatory elements in human cardiomyocytes S. Haydar et al. Nature Communications 2026 Jan

Abstract

Cis-regulatory elements (CREs) are noncoding DNA regions regulating cell-type-specific gene expression programs by interacting with distal gene promoters. Here, we aim to decode the function and spatial organization of CRE-promoter interactions in human cardiomyocytes. We analyzed the epigenome and chromatin interactions of human male atrial, ventricular, and failing cardiomyocytes. Atrial and ventricular cardiomyocytes harbored chamber-specific CRE-promoter interactions modulating gene expression as confirmed by functional epigenetic silencing. These CRE-promoter interactions explain the distinct contribution of non-coding genetic variants to atrial and ventricular diseases, such as dilated cardiomyopathy and arrhythmias. We dissected the prototypic KCNJ2 locus, encoding a potassium channel associated with ventricular arrhythmia susceptibility. Functional epigenetic silencing confirmed that CREs, harboring QT-duration-associated genetic risk factors, modulate KCNJ2 gene expression levels, alter KCNJ2-dependent channel currents, and affect cardiomyocyte repolarization. The presented human CM-specific chromatin interaction analysis provides key insights into regulatory mechanisms and aids in interpreting genetic risk factors. Here the authors functionally test and resolve the spatial genome organization of cis-regulatory elements and genetic variants in atrial, ventricular, and failing human cardiomyocytes and linked them to heart disease traits, including QT syndrome.