# The score describes amounts of databases that predict the corresponding gene target of a miRNA out of 12 selected database in miRWalk.
These targets were examined further below.

KO/KD cell lines

In order to test the effect of reduced/loss-of-function miRNAs for three specific miRNAs – miR-128, miR-34C, and miR-181, which all have potential targets in glycosylation – and evaluate possible correlation between selected miRNAs and N-glycosylation, we set up a CRISPR-based engineering strategy.
For each target, two plasmids were generated, each expressing one specific guide RNA (gRNA) targeting the upstream and downstream region of the miRNA expression site in the CHO genome (up-sgRNA and dw-sgRNA respectively).
For each target, up-sgRNA and dw-sgRNA plasmids were co-transfected into CHO-S cells, and subsequently single cell sorting and screened. As shown in Figure 2A , KO/KD of miRNA was first verified at the molecular level by genomic DNA PCR. It was possible to isolate both miR-128, miR-34C, and miR-181 KO/KD clones which showed a clean deletion (KO). For some miR-128 and miR-34c clonal cell lines, both wt bands and KO bands were found in the gel. This indicates that the deletion may only have happened in one allele in these cell lines, thus making them KD rather than KO, and suggesting that these exist in more than one allele.