Volume 7 Issue 5, May 2023

Prime editing can efficiently rewrite the genetic mutation causing sickle cell disease, in haematopoietic stem cells from patients.

Amphiphilic peptides can aid the delivery of CRISPR ribonucleoproteins into primary human lymphocytes at low toxicity, boosting editing yields with respect to the use of electroporation.

Each of the 13 protein-coding genes in the mouse mitochondrial genome can be ablated using a library of optimized double-stranded-DNA deaminase-derived cytosine base editors.

We engineered integrase-deficient lentiviruses to act as vectors for the delivery of large gene knock-ins via homology-directed repair. This technology enables the non-cytotoxic, targeted insertion of difficult-to-express transgenes into genomic loci that are essential to cell survival, thereby overcoming the gene silencing that otherwise limits primary immune cell engineering.

The inability to precisely manipulate mammalian mitochondrial DNA has stalled our understanding of mitochondrial biology and the generation of cellular and animal models in which to study it. DNA base editing technologies have enabled the generation of a library of mitochondrial base editors that precisely ablate every protein-coding gene in the mouse mitochondrial genome.

Prime editing can efficiently correct the sickle-cell allele to produce wild-type haemoglobin in patient haematopoietic stem cells that engraft efficiently in mice, yielding erythrocytes resistant to hypoxia-induced sickling.

A Cas9-based gene therapy that replaces expanded CAG repeats in the mutant HTT allele causing Huntington’s disease by a normal CAG repeat led to substantial reductions in neurological symptoms in a pig model of the disease.

The yields of edited primary human lymphocytes can be increased substantially, with respect to those obtained via electroporation, by delivering a CRISPR ribonucleoprotein alongside an amphiphilic peptide identified via screening.

A method leveraging an integrase-deficient lentivirus, homology-directed repair and the electroporation of a CRISPR-associated ribonucleoprotein complex allows for the knock-in and stable expression of large payloads in primary human cells.

The activity of standard Cas9-based genome-editing systems can be constrained by the addition of cytosine stretches to the 5′-end of conventional single-guide RNAs.

A library of double-stranded-DNA deaminase-derived cytosine base editors allows for the precise ablation of every mtDNA protein-coding gene in the mouse mitochondrial genome.