Fig. 5: Testing compatibility of near PAM less spG Cas9 in NT-CRISPR. | Communications Biology

Fig. 5: Testing compatibility of near PAM less spG Cas9 in NT-CRISPR.

From: NT-CRISPR, combining natural transformation and CRISPR-Cas9 counterselection for markerless and scarless genome editing in Vibrio natriegens

Fig. 5: Testing compatibility of near PAM less spG Cas9 in NT-CRISPR.

a Killing efficiency with spG Cas9 with all possible PAM sequences. Cas9 with NGG PAM is shown as a reference in gray. Killing efficiency is calculated as follows: \({{{{{\mathrm{Killing}}}}}}\,{{{{{\rm{efficiency}}}}}}[ \% ]=1-\frac{{{{{{\rm{CFU}}}}}}/\mu {{{{{\rm{L}}}}}}\,{{{{{\rm{with}}}}}}\,{{{{{\rm{counterselection}}}}}}}{{{{{{\rm{CFU}}}}}}/\mu {{{{{\rm{L}}}}}}\,{{{{{\rm{without}}}}}}\,{{{{{\rm{counterselection}}}}}}}\,\ast 100\). n = 4 replicates, representing two independent biological replicates (circle or triangle) and two independent experiments (filled or open symbols). Bars show the mean of all replicates and error bars indicate standard deviation of the mean. The dashed line indicates the highest possible value. b Efficiency for integration of mScarlet-I using either SpG Cas9 with a NGT PAM sequence or using the indirect approach described above. Integration of mScarlet-I was identified by PCR assays (n = 50 colonies).

Back to article page