A protocol for custom CRISPR Cas9 donor vector construction to truncate genes in mammalian cells using pcDNA3 backbone

Table 1 PCR Primers utilized to amplify FOXO3 gene fragments for Donor Vector Gibson Assembly Reactions

Primer	Sequence^a
FOXO3 ARM 1 F	5′-GTGCTTTACGGCACCTCGACCCCCCGGCACAACCTGTCACTGC-3′
FOXO3 ARM 1 R	5′-CCGTCTATCAGGGCGATGGCCGCTGTAGAGCATGGGCGAGAG-3′
FOXO3 ARM 2 F	5′-CAAATAAAGCATTTTTTTCACTCGGTGGAACTGCCACGGCTG-3′
FOXO3 ARM 2 R	5′-GAGTTTGGACAAACCACAACTAGGTCCAAGTCGCTGGGGAAC-3′

^aNucleotides in bold corresponds to pcDNA3 vector sequences. Sequences in italics were to amplify the indicated FOXO3 arm

ISSN: 1471-2199