CRISPR-activation mediated long non-coding RNA overexpression in Komagataella phaffii

Here RNA-seq was used to examine gene expression differences when targeting long non-coding RNA (lncRNA) overexpression in a CRISPR-activation mediated manner in Komagataella phaffii. Strains were sequenced after small-scale, glucose-limited fed-batch like screenings of recombinant protein producing strains. A number of strains were used, including a strain without the CRISPR activation machinery, a no-targeting control that expressed all components including a scaffold-guide RNA (scgRNAs) not targeted to the K. phaffii genome, and other strains expressing scgRNAs targeted near the predicted transcript start site of lncRNAs.