CRISPR/Cas9 mediated editing of the Quorn fungus Fusarium venenatum A3/5 by transient expression of Cas9 and sgRNAs targeting endogenous marker gene PKS12

Table 2 Viability of F. venenatum colonies after transformation with AMA1 CRISPR vectors

Vector	A	B	C
pFCFvCas9	3	0	3 (0)
pFC332	3	2	1 (0)
pFCFvCas9::PolII-PK3/14	1	0	1 (0)
pFC332::5S-PK3/14	12	7	5 (0)
pFC332::PolII-PK3/14	27	25	2 (2)

Viability of colonies from protoplasts transformed with empty CRISPR vectors pFCFvCas9 or pFC332 (expressing cas9 codon optimised for F. venenatum (Fv) or A. niger), and vectors with single sgRNA constructs transcribed from the Fv5SrRNA promoter (pFC332::5S-PK3/14) or the PgdpA-dual ribozyme cassette (pFCFvCas9::PolII-PK3/14 and pFC332::PolII-PK3/14). Column A: numbers of colonies transferred to hygromycin selective medium; Column B: numbers of colonies showing growth after 14 days culture on selective medium; Column C: numbers of colonies not able to grow on selective medium (of these, numbers that subsequently showed growth after transfer to non-selective medium).

ISSN: 2054-3085