Effects of IL2RA non-coding and coding sequence edits on IL2RA expression in primary human T cells.. Homo sapiens

We edited an intronic IL2RA enhancer or exon in primary human naïve T cells. Electroporated cells were activated and sorted on IL2RA expression over three days. Amplicon sequencing was used to assess the effects on sequence edits on IL2RA expression. Overall design: Cas9 ribonucleoproteins were targeted to the intronic IL2RA enhancer (SNP or CaRE4-Mix) or exon in primary human naïve T cells. Edited cells were activated with anti-CD3 and anti-CD28 antibodies and genomic DNA was extracted from unsorted, IL2RA- or IL2RA+ cells over three days. Amplicon sequencing across the cut sites was performed to assess the sequence edits in the sorted populations.