Construction of BEAS-2B cell line with NLRC5 overexpression through PiggyBac transposon and its function study
SUN Danni
LIU Xinyue
RAO Zifan
DU Lan
ZHANG Xianglin
LIU Ziyu
HOU Shiyuan
SHEN Xing
LIU Rongrong
WU Xing'an
Abstract：Objective To construct a BEAS-2B cell line stably overexpressing NLRC5 by using PiggyBac transposon system,so as to provide a powerful tool for further exploration of viral antigen presentation mechanisms.Methods Based on NLRC5 gene data,the protein-coding sequence of NLRC5 was amplified,and a transposon plasmid containing NLRC5 was constructed.The helper plasmid and transposon plasmid were co-transfected into BEAS-2B cells.Following puromycin screening for stably transduced cells,monoclonalization was conducted to validate molecular-level overexpression in the obtained monoclonal cells.Results A BEAS-2B cell line,which overexpressed NLRC5,was successfully constructed while maintaining regular cell viability and proliferation capacity.Conclusion The overexpression of NLRC5 is accomplished using PiggyBac transposon,resulting in heightened expression of MHC-Ⅰ molecule genes.Investigation into the regulatory role of NLRC5 in the MHC-Ⅰ antigen peptide presentation pathway via its overexpression offers substantial insights into the intricate mechanisms of antiviral immunity,thus facilitating the development of novel therapeutic approaches.
Keywords：NLRC5PiggyBac transposongene overexpressionstably transfected cell line
Publication Date：2025-04-30
Online Publishing Date：2026-08-26(First online date of this platform, not the publication date of the document)
Pages：7( 526-532 )