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.
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