Journal: Journal of Cellular and Molecular Medicine
Article Title: Deregulated lncRNA expression profile in the mouse lung adenocarcinomas with KRAS‐G12D mutation and P53 knockout
doi: 10.1111/jcmm.14584
Figure Lengend Snippet: Construction of KP cell lines with Kras gene disruption and identification of Kras related lncRNAs in KP cells. A, Messy peak figure reporting the results of DNA sequencing showed mutations induced by Cas9/gRNA in the Kras guide RNA. The stable Cas9 expression KP cells (KP‐Cas9) were developed by pCDH‐bla‐Cas9 lentivirus infection and selection with 10 μg/mL blasticidin. KP‐Cas9 cells were further infected by the Kras ‐targeting guide RNA expression lentivirus and selected by 4 μg/mL puromycin, lenti‐guide empty vector lentivirus as negative control. The DNA fragment containing the Kras guide RNA binding site was amplified by using PCR with the genomic DNA of guide RNA‐infected KP‐cas9 cells. B, Comparison between microarray data and qPCR results of lncRNAs regulated by Kras . Among the selected 210 lncRNAs, qPCR results determined 33 lncRNAs, whose expression was altered by the Kras deletion in KP cells and exhibited opposite expression alteration direction to that in the microarray of the mouse lung adenocarcinoma tissues. The heights of the columns in the chart represent the log‐transformed median fold changes (tumour sample/normal tissues in microarray data of the mouse lung adenocarcinoma tissues or KP‐Kras‐guide cells/ KP‐guide‐NC cells in qPCR results). Mean ± SD are shown, n = 3 or 2
Article Snippet: For microarray analysis, lncRNA + mRNA mouse Gene Expression Microarray v1.0, 4x180K chip (CapitalBio Corp), was employed and conducted by CapitalBio Corp according to the manufacturer's protocols.
Techniques: Disruption, DNA Sequencing, Expressing, Infection, Selection, RNA Expression, Plasmid Preparation, Negative Control, RNA Binding Assay, Amplification, Comparison, Microarray, Transformation Assay