Data from: Engineering self-deliverable ribonucleoproteins for genome editing in the brain

The delivery of CRISPR ribonucleoproteins (RNPs) for genome editing in
vitro and in vivo has important advantages over other delivery methods,
including reduced off-target and immunogenic effects. However, effective
delivery of RNPs remains challenging in certain cell types due to low
efficiency and cell toxicity. To address these issues, we engineered
self-deliverable RNPs that can promote efficient cellular uptake and carry
out robust genome editing without the need for helper materials or
biomolecules. Screening of cell-penetrating peptides (CPPs) fused to
CRISPR-Cas9 protein identified potent constructs capable of efficient
genome editing of neural progenitor cells. Further engineering of these
fusion proteins identified a C-terminal Cas9 fusion with three copies of
A22p, a peptide derived from human semaphorin-3a, that exhibited
substantially improved editing efficacy compared to other constructs. We
found that self-deliverable Cas9 RNPs generated robust genome edits in
clinically relevant genes when injected directly into the mouse striatum.
Overall, self-deliverable Cas9 proteins provide a facile and effective
platform for genome editing in vitro and in vivo.