Fig. 2.

Fig. 2. Simultaneous editing of galK and xylB targets. (A) sgRNAs recognizing galK and xylB target sequences. Mutations are introduced through oligonucleotides, resulting in changes from ⁵⁰⁴TAAC to ATCA for the galK target and from ⁶⁴⁹GCGA to AACT for the xylB target. Each sgRNA possesses a 20 nt long TRS specific to the galK or xylB target. Gray boxes indicate PAM sequences and target nucleotides are marked in red. (B) Optimization of recovery temperature for multiplex genome editing. Recovery temperatures of 37, 27, and 17°C were investigated, with varying recovery times at each temperature. Gray and purple bars indicate editing efficiency and the number of surviving cells, respectively.

© J. Microbiol. Biotechnol.