Fig. 2

A LINC00511 gene schematic, gene length, sgRNA binding site, and gene primers. The position of the fragment deleted by the CRISPR / Cas9 technique is depicted in orange in this diagram. B expression portion of sgRNA in vector PX459. The BbsI enzyme is used to form a sticky end, which is then supplemented by sgRNA tags. C sgRNAs 1 and 2 results in the Px459 vector. No. 1 is a 100 bp DNA ladder. Nos. 2 and 3 PCR reactions were performed on recombinant vectors PX459-sgRNA1 and PX459-sgRNA2, respectively, yielding 276 bp bands. Negative PCR control (no. 4) (without DNA). D Column 1: Heterozygous cells with only one LINC00511 gene allele degraded and two bands 1996 and 480 visible. 2: Control group (PX459-control), in which just one band of 1996 pairs are transfected and the blank vector is transfected. This value indicates that the LINC00511 gene hasn't been degraded. Column 3 100-bp marker. Column 4: No vectors have been transfected blank control cells, and the 1996 band shows the pair. Column 5: Cells that have lost both alleles of the target gene and contain a small band of roughly 480 kb. Column 6: Negative control (no DNA in any of the PCR reagents). E LINC00511 transcript levels in MCF-HGH and MDA-MB-468 breast cancer cells compared to GAPDH levels in wild-type and "CRISPR " knockout cells