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| Product Name | NLS-Cas9-EGFP Nuclease with C-Terminal EGFP and 6xHis Tags |
| Catalog No. | IVDRM-0509 |
| Description | An engineered Cas9 nuclease carrying an N-terminal nuclear localisation signal together with a C-terminal EGFP reporter and a 6xHis sequence. Delivered as a guide RNA directed RNP complex, for example by electroporation, the protein localises immediately to the nucleus on entry, avoiding the delay of in vivo transcription or translation. The EGFP tag acts as a reporter for tracking transfected cells and enables enrichment of the genome-edited population by fluorescence activated cell sorting, which greatly reduces the labour and cost associated with single cell cloning and genotyping. In vitro cleavage of linearised plasmid substrate exceeds 90% digestion efficiency in a 20 uL reaction with Cas9 nuclease buffer at 37°C for one hour. |
| Intended Use | Genome editing, cell transfection studies and FACS enrichment of edited cell populations |
| Principle / Technology | Guide RNA directed Cas9 cleavage with EGFP fluorescence reporting transfected cells for sorting |
| Sample Type | Cultured cells transfected with the Cas9 RNP complex |
| Specificity | N-terminal NLS, C-terminal EGFP and 6xHis tag; linearised plasmid digestion efficiency above 90% |
| Reaction Conditions / Protocol | Combine with guide RNA, deliver the RNP by electroporation, then sort EGFP positive cells by FACS |
| Storage Conditions | -25 to -15°C |
| Shelf Life | 1 year |
| Package Specifications | 50 ug; 100 ug |
| Key Features | Nuclear localisation on cell entry; fluorescent reporter for tracking and sorting edited cells |
| Readouts / Downstream Assays | Fluorescence microscopy, FACS profiles and cleavage efficiency on target substrate |
For in vitro diagnostic use.
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