aRGEN - active RGEN- i.e. CRISPR system whereas the sgRNA is in RNA format and the Cas9 is delivered as recombinant, active protein.
RNP Editing Tool
Highly efficient RNA-guided genome editing system via delivery of purified Cas9 ribonucleoproteins
RNA-guided engineered nucleases (RGENs) derived from the prokaryotic adaptive immune system known as CRISPR (clustered, regularly interspaced, short palindromic repeat)/Cas (CRISPR-associated) enable genome editing in human cell lines, animals, and plants but are limited by off-target effects and unwanted integration of DNA segments derived from plasmids encoding Cas9 and guide RNA at both on-target and off-target sites in the genome.
RNPs ribonucleoproteins i.e. purified recombinant Cas9 protein and guide RNA can overcome these limitations. RNPs delivered into cultured human cells, including hard-totransfect fibroblasts and pluripotent stem cells induce site-specific mutations at frequencies of up to 79%, while reducing off-target mutations associated with plasmid transfection at off-target sites that differ only by one or two nucleotides from on-target sites
RNPs cleave chromosomal DNA almost immediately after delivery and are degraded rapidly in cells, reducing off-target effects. Furthermore, RNP delivery is less stressful to human embryonic stem cells, producing at least two-fold more colonies than does plasmid transfection
1. Simple procedure - aRGEN can be directly delivered into cell line and show excellent activity
(Tested with nucleofection by Amaxa nucleofector and Neon mediated electroporation)
2. Good for cell lines where DNA delivery is less efficient
(Tested on primary fibroblasts and H9 human ES cells which are relatively harder to transfect than common cell lines)
3. KO/KI/chromosomal deletion all possible
4. Improved specificity
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