Table 1 Primers for LDLR Sanger sequencing and targeted insertion used in this study
Gene | Forward (5′–3′) | Reverse (5′–3′) | Annealing temp. (°C) |
|---|---|---|---|
Primers used to screen FH mutation | |||
LDLR (exon 2) | TTTCCAGCTACGACACAGCAGGT | AGAACTGAGCAATCAAGCGGTTGA | 55.8 |
Primers used for assessment of the targeted integration of the LDLR cassette | |||
5′ junction | CTGCCGTCTCTCTCCTGAGT (5′ arm) | GTGGGCTTGTACTCGGTCAT (Puro(R)) | 62 |
3′ junction | GTCAGCTCCTTTCCGGGACT (WPRE) | GGAACGGGGCTCAGTCTG (3′ arm) | 62 |
AAVS1 targeted site by CRISPR/CAS9 | GCCCTATGTCCACTTCAGGA | ACAGGAGGTGGGGGTTAGAC | 62 |