Chapter 5 —— 142 —— Map and nucleotide sequence of selector AAV donor construct BI19_pAAV-HRLMN.A1. Diagram of AAV-HRLMN.A1 vector genome containing an LMNA-targeting HR donor template (1762 bp). The selector donor DNA sequence (964 bp) is designed for installing the ouabain resistance gain-of-function Q118R and N129D SNPs (RD) within the exon 4 of ATP1A1 upon gRNA GA1-directed cleavage within intron 4. AAV ITR, adeno-associated virus type2 inverted terminal repeat; DNA sequences homologous to target LMNA alleles (homology arms) flank the mScarlet-1 reporter coding sequence (red arrow). The gRNALMNA directs HR-mediated LMNA gene tagging upon targeted DNA cleavage at the N-terminus of LMNA alleles. U6 promoter, human U6 snRNA polymerase III promoter driving the expression of each gRNA. The vector plasmid backbone is not shown. ctccatttctgacccttcctgtgtggtctttagaagggaattggatcctagagtccggccgcccccttcaccga gggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatt tgactgtaaacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagtt ttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagtatttcgatttcttggctttata tatcttgtggaaaggacgaaacaccgagttctgtaattcagcatagtttcagagctatgctggaaacagcatag caagttgaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgctttttttgaattgatcc atatatagggcccgggttataattacctcaggtcgacgtcccatgtgcaggtgctgaattgtagataagtagca tggcgggttaatcattaactacaaggaacccctagtgatggagttggccactccctctctgcgcgctcgctcgc tcactgaggccgggcgaccaaaggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcg cgcag
RkJQdWJsaXNoZXIy MTk4NDMw