Improved specificity and efficacy of base-editing therapies with hybrid guide RNAs.

Phenylketonuria (PKU), hereditary tyrosinemia type 1 (HT1), and mucopolysaccharidosis type 1 (MPSI) are autosomal recessive disorders linked to the phenylalanine hydroxylase () gene, fumarylacetoacetate hydrolase () gene, and alpha-L-iduronidase () gene, respectively. Potential therapeutic strategies to ameliorate disease include corrective editing of pathogenic variants in the and genes and, as a variant-agnostic approach, inactivation of the 4-hydroxyphenylpyruvate dioxygenase () gene, a modifier of HT1, via adenine base editing. Here we evaluated the off-target editing profiles of therapeutic lead guide RNAs (gRNAs) that, when combined with adenine base editors correct the recurrent P281L variant, R408W variant, or W402X variant or disrupt the gene in human hepatocytes.

IVT generation of guideRNAs for Cas9-enrichment Nanopore Sequencing.

Generating high-coverage sequencing coverage at select genomic loci has extensive applications in both research science and genetic medicine. Long-read sequencing technologies (e.g.