CRISPR/Cas9 delivery methods
For gene editing in mammalian cells we typically recommend delivery of a ribonuclear protein (RNP) complex consisting of Cas9 protein and a single guide RNA (sgRNA). RNPs remain in the cell for a short time and the dose is minimal, leading to lower toxicity and reduced editing at off-target sites compared to other methods.
RNPs can be delivered using either electroporation of recombinant Cas9 protein along with in vitro transcribed guide RNA or using cell-derived nanovesicles called gesicles. We also provide kits for delivery using lentivirus, AAV, and plasmid. See the links below for articles and tech notes related to delivery of CRISPR/Cas9 elements.
Delivery of RNPs using electroporation
Electroporation-grade Cas9
Our Cas9 performs highly efficient gene editing, including in iPS and hematopoietic stem cells.
In vitro transcription of sgRNA
Kits providing a simple method for production and evaluation of sgRNAs for Cas9-mediated gene editing.
Knockins in iPS cells using RNPs and ssDNA
Data demonstrating efficient genome editing of iPS cells using HDR templates generated with the Guide-it Long ssDNA Production System.
Delivery of RNPs using gesicles
Gesicle technology overview
Learn about Guide-it CRISPR/Cas9 Gesicle Production System components and workflow.
Gesicles function in a broad range of cells
Gesicles containing custom sgRNA/Cas9 ribonucleoprotein complexes can be made and used to achieve genome editing in a broad range of cell types.
Gesicles reduce off-target effects
This tech note discusses using CRISPR/Cas9 Gesicles for genome editing while avoiding unwanted off-target effects.
CRISPR-mediated editing using virus or plasmid
CRISPR/Cas9 gene editing with AAV
Use viral transduction to overcome difficulties with plasmid transfection delivery of Cas9 and sgRNA to many mammalian cell types.
Tet-inducible Cas9
This system enables lentiviral delivery of the components necessary for doxycycline-induced Cas9 expression.
Plasmid systems
Kits for cloning and expression of single guide RNAs for mammalian genome editing using CRISPR/Cas9 technology.
Lentiviral systems
The Lenti-X and Lenti-X Tet-On 3G CRISPR/Cas9 systems are complete systems for lentiviral-mediated CRISPR/Cas9 genome editing.
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