Efficient multiplex mutagenesis by RNA-guided Cas9 and its use in the characterization of regulatory elements in the AGAMOUS gene

Table 1 Efficiency of multiplex editing by using different Cas9 expression cassettes

Target gene (s) (construct)	# Plants	elf6	ref6	at5g46910	Triple
Target gene (s) (construct)	# Plants	# Plants	# Plants	# Plants	# Plants
ELF6/REF6/At5g46910 (p35S::hucoCas9)	48	0	0	0	0
ELF6/REF6/At5g46910 (p35S::pcoCas9)	41	3	2	0	0
ELF6/REF6/At5g46910 (pUBQ10::hucoCas9)	48	14	12	7	6
ELF6/REF6/At5g46910 (pUBQ10::pcoCas9)	48	5	22	12	5

Leaf samples from T1 transformants were genotyped by PCR. Mutation was scored when a band with expected deletion was obtained. Triple-mutation (triple) means that with the same plant, mutated bands for all the three loci could be detected. #, number of

ISSN: 1746-4811