ArciTect™ T7 Endonuclease I Kit

Complete kit for optimization of genome editing in human cells

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The ArciTect™ Human CRISPR Optimization Kit is a flow cytometry-based kit designed to enable rapid optimization of genome editing in human cells. It can also be used as a positive control for experiments using the ArciTect™ CRISPR-Cas9 genome editing system in human cells. The kit comprises ArciTect™ Human B2M sgRNA, ArciTect™ Cas9 Nuclease, and a fluorophore-conjugated beta-2 microglobulin (B2M) antibody. ArciTect™ Cas9 Nuclease must first be complexed with ArciTect™ Human B2M sgRNA, followed by delivery into human cells and subsequent culture for 48 - 96 hours. Following culture, editing efficiency can be assessed quickly by collection and processing for flow cytometry using a fluorophore-conjugated B2M antibody.

This kit has been tested and validated for use with the ArciTect™ line of genome editing products. By targeting the B2M gene that encodes a ubiquitously expressed cell-surface protein, editing efficiency can be rapidly and quantitatively assessed using flow cytometry methods. This technique also enables additional evaluation of editing success such as viability monitoring and/or assessment of cell type-specific marker expression

Data Figures

Figure 1. ArciTect™ Human CRISPR Optimization Kit Exhibits High Editing Efficiency in hPSCs, T Cells, and CD34+ Hematopoietic Stem and Progenitor Cells (HSPCs)

Cells were electroporated with RNP complexes containing sgRNA targeting B2M complexed with ArciTect™ Cas9 Nuclease protein. The cells were cultured for 72 hours following electroporation and editing efficiency was monitored by flow cytometry to detect loss of B2M expression; n = 3 biological replicates (WLS-1C iPSCs/hPSCs) or n = 3 donors (T cells and CD34+ HSPCs). No EP: non-electroporated controls.

Figure 2. Anti-Human Beta-2-Microglobulin, Clone #35 Conjugates Exhibit Equivalent Performance Across Samples to Monitor B2M Knockout Efficiency Using the ArciTect™ Human CRISPR Optimization Kit

CD34+ HSPCs isolated from two independent donors were electroporated with RNP complexes containing sgRNA targeting B2M complexed with ArciTect™ Cas9 Nuclease protein. The cells were cultured for 96 hours following electroporation and editing efficiency was monitored by flow cytometry to detect loss of B2M expression using anti-human beta-2-microglobulin, clone #35, (A, B) APC-conjugated, (C, D) FITC-conjugated, and (E, F) PE-conjugated. The histograms were derived from gated events of viable (7-AAD-negative) cells. Orange histograms: RNP-electroporated samples; grey histograms: non-electroporated control samples.

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
Catalog #
100-0470, 100-0472, 100-0471
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
100-0470
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
100-0470, 100-0472, 100-0471
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
100-0470, 100-0472, 100-0471
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
100-0472
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
100-0471
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.

Legal Statement: The purchase of the ArciTect™ products conveys to the purchaser the limited, non-transferable right to use, in accordance with all applicable laws and regulations, the ArciTect™ products and any related material solely for research purposes only, not for any clinical, human, agricultural, veterinary, livestock, or commercial purpose, with no warranty (express or implied) from the owners or assignees of the Patents or STEMCELL Technologies each of whom expressly disclaim any warranty regarding results obtained through the use of the ArciTect™ products, and without any exception the owners or assignees of the Patents or STEMCELL Technologies, or as applicable a director, trustee, officer, employee, agent, faculty, official investigator or student thereof, will not suffer or be exposed to any liability, damages, loss, or expense of any kind whatsoever arising from or related to the use of the ArciTect™ products by a purchaser of same. Distribution of ArciTect™ products by STEMCELL Technologies is covered under at least US 8,697,359, US 8,771,945, US 8,795,965, US 8,865,406, US 8,871,445, US 8,889,356, US 8,889,418, US 8,895,308, US 8,906,616 and foreign equivalents (“Patents”). PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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