## CRISPR/Cas9-based mutagenesis of GBP homologues in barley
The CRISPOR web tool80 (version 4.97) was used to design two single guide RNAs for each *GBP1* and *GBP2*:
GBP1_gRNA1: 5’-CCCGGCACGCTTCTTCGCGCCGG-3’
GBP1_gRNA2: 5’-TGGCGCCTTCGGATGAACAGCGG-3’
GBP2_gRNA1: 5’-TAAGATCCGTCGAGGCAGTATGG-3’
GBP2_gRNA2: 5’-GTACAGCCGTTGCTACCCGACGG-3’
Golden Gate cloning was used to load each guide sequence from complementary oligonucleotides into shuttle vectors pMGE625, pMGE626, pMGE628, pMGE629. The four guide expression cassettes were then assembled into binary vector pMGE599 to create pMP202. Vectors and cloning protocols have been previously described87 and were kindly provided by Johannes Stuttmann. Stable transformation of pMP202 in cv. Golden Promise Fast was performed as previously described.88
