Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain
the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in
Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles
and JavaScript.
Stable transposon-driven integration of transgenes into immune cells
This issue highlights optimized base editors targeting the exon-7 mutation in SMN2 to restore the expression of the survival motor neuron protein to normal levels, the virus-mediated delivery of a transposon and an mRNA-encoded transposase for the integration of transgenes into immune cells, the enhanced viability of genome-engineered T cells transfected by electroporation via an isotonic buffer that dampens cytosolic cGAS–DNA interactions, the annotation of variants of the BRCA2 gene in human pluripotent stem cells, that host genes involved in viral processes can constrain the lentiviral delivery and expression of Cas13, and that replacing amino acid residues in an immunodominant and conserved T cell epitope in the capsid of an adeno-associated virus can abrogate its immunogenicity while preserving its function and potency.
The cover illustrates a gene-delivery system that enhances the stability of the integration of a desired transgene in immune cells by relying on a Sleeping Beauty transposase encoded into an mRNA delivered by an adeno-associated virus.
Appealing against an editor’s negative decision is likely to be more fruitful when considering the basis of the editorial assessment and offering ways forward.
Base editors can restore the expression of survival motor neuron protein to therapeutically beneficial levels in animal and cell models of spinal muscular atrophy.
We functionally assessed clinically observed mutations of the BRCA2 gene and analysed structure–function relationships of variants of the gene by using high-throughput CRISPR-mediated mutagenesis and pooled screening in locally haploid human pluripotent stem cells and in fibroblasts differentiated from them.
We revealed that the RNA-targeting activity of the Cas13 family of nucleases allows them to directly target endogenous RNA in mammalian cells. Such activity limits the usage of lentiviral Cas13 systems, and suggests a need for caution when applying Cas13-based systems.
Optimized base editors targeting the exon-7 mutation in SMN2 restore expression of the survival motor neuron (SMN) protein to normal levels, as shown in mice with spinal muscular atrophy and in fibroblasts from patients with this genetic disease.
A gene-delivery system relying on a Sleeping Beauty transposase encoded into an mRNA delivered by an adeno-associated virus enhances the stability and efficiency of the integration and expression of a desired transgene in immune cells.
Human pluripotent stem cells with one copy of the BRCA2 gene deleted can be used to annotate variants of the gene and to test their sensitivities to inhibition by the poly(ADP-ribose) polymerase.
Cas13 can intrinsically target host RNA in mammalian cells through previously unrecognized mechanisms without the involvement of a CRISPR RNA, and host genes involved in viral processes can constrain the lentiviral delivery and expression of Cas13.
Replacing amino acid residues in an immunodominant and conserved T-cell epitope in the capsid of an adeno-associated virus widely used in gene therapies abrogates its immunogenicity while preserving its function and potency.