CRISPR-Cas9 targeted-knockout human colorectal cancer cell PPP1R1C gene and specific sgRNA thereof

Abstract

The invention discloses a CRISPR-Cas9 targeted-knockout human colorectal cancer cell PPP1R1C gene and a specific sgRNA sequence. The invention comprises the following steps: firstly, acquiring sgRNA of a specific targeting PPP1R1C gene functional area, wherein a base sequence of the sgRNA is shown as SEQ ID NO.1; then, constructing an sgRNA-lentiviral vector system of the PPP1RQC gene, wherein thesystem contains Cas9 protein; and finally, infecting human colorectal cancer HT-29 cells with CRISPR/Cas9 lentivirus containing the sgRNA, so that a cell line that a PPP1R1C protein expression levelis obviously reduced is obtained. The invention is simple in operating steps, good in sgRNA targeting performance and high in cutting efficiency on the PPP1R1C gene; the constructed CRISPR/Cas9 lentiviral system is high in knockout efficiency and is capable of achieving specific knockout of the PPP1R1C gene, so that the human colorectal cancer cells that the PPP1R1C gene is knocked out is obtained; therefore, a powerful tool is provided for further researching an action mechanism of the PPP1R1C in the colorectal cancer cells.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and more specifically relates to CRISPR / Cas9 targeted knockout of human intestinal cancer cell PPP1R1C gene and its specific gRNA. Background technique [0002] Protein phosphatase 1 (PP1) has serine / threonine phosphatase activity and is widely involved in cell signal transduction processes. Protein kinases and protein phosphatases can phosphorylate and dephosphorylate a variety of proteins, thereby changing the charge, solubility, and binding capacity of enzymes, receptors, and ion channels. They are widely involved in cell division, gene expression regulation, and neural information transmission. , muscle contraction and cell signal transduction and other important life processes. Impaired activity of protein phosphatases PP1 and PP2A can lead to cell dysfunction, such as tumors and abnormal neuronal differentiation. Protein phosphatase 1 regulatory inhibitor subunit 1C (protein phosphatase...

AI Technical Summary

Problems solved by technology

At present, researchers have done a lot of research on the pathogenesis of colorectal cancer, and have discovered many pathways and mechanisms involved in the progression of colorectal cancer, such as activation of proto-oncogenes, inactivation of tumor suppressor genes (point mutations, rearrangements, deletions) and chromosomal Abnormalities, etc., but the molecular mechanisms involved in the occurrence and progression of colorectal cancer are still unclear, and further research is needed

Images