Year: 2019
Type: article
Abstract: Techniques for exclusion of exons from mature transcripts have been applied as gene therapies for treating many different diseases. Since exon skipping has been traditionally accomplished using technologies that have a transient effect, it is particularly important to develop new techniques that enable permanent exon skipping. We have recently shown that this can be accomplished using cytidine base editors for permanently disabling the splice acceptor of target exons. We now demonstrate the application of CRISPR-Cas9 adenine deaminase base editors to disrupt the conserved adenine within splice acceptor sites for programmable exon skipping. We also demonstrate that by altering the amino acid sequence of the linker between the adenosine deaminase domain and the Cas9-nickase or by coupling the adenine base editor with a uracil glycosylase inhibitor, the DNA editing efficiency and exon-skipping rates improve significantly. Finally, we developed a split base editor architecture compatible with adeno-associated viral packaging. Collectively, these results represent significant progress toward permanent in vivo exon skipping through base editing and, ultimately, a new modality of gene therapy for the treatment of genetic diseases.
Source: Cell Discovery
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,100
Awards: G1336039831, G1455788304, G1714404606, G2338432649, CORRECTION OF DYSTROPHIN CARDIOMYOPATHIES BY ACTIVATION OF NATIVE UTRN EXPRESSION, Engineering platforms for genome-scale screening of transcriptional activation domains, PFC: Center for the Physics of Living Cells, G5866859489, Graduate Research Fellowship Program (GRFP), G6638377572, G9000033857, Predicting Transcriptional and Epigenetic Networks in Cancer from Sequencing Data, Engineering platforms for editing RNA with single base resolution
OpenAlex ID: W2969635671