Journal: American journal of human genetics

Article Title: Genetic and functional correction of argininosuccinate lyase deficiency using CRISPR adenine base editors.

doi: 10.1016/j.ajhg.2024.03.004

Figure Lengend Snippet: Figure 3. Editing efficiency, toxicity profile, and rescue of the metabolic phenotype in primary fibroblasts after lipid nanoparticles ABE8e treatment (A) Diagram of the lipid nanoparticle contents: an sgRNA targeting the ASL c.1153C>T variant plus an RNA cassette for ABE8e expres- sion. The spacer section of the sgRNA, which targets the DNA, is written from base 1 to base 20. The PAM, not included in the sgRNA oligo, corresponds to bases 21–23 (GGG). (B) On target A-to-G editing efficiency. We employed primary fibroblasts from two different individuals. We independently treated these fibroblasts in duplicates (four biological replicates) with eight different doses (0–5,100 ng RNA) of three types of lipid nanoparticle ABE8e (legend continued on next page)

Article Snippet: In vitro transcription ABEmax, and ABE8e Employing the ABEmax or the ABE8e IVT plasmid (Addgene: 201676 and 171761) as a DNA template, T3 RNA transcription was performed according to the manufacturer’s protocol (mMESSAGE mMACHINE T3 Transcription Kit, Thermo Fisher Scientific, Invitrogen, catalog no.: AM1348).

Techniques: Variant Assay