NDLI: Spatiotemporal control of porcine p65RHD expression by advanced Tet-On system in PIEC cells helps regulate NFкB activity

Content Provider	Springer Nature Link
Author	Ruan, Jinxue Liu, Nan Ouyang, Hongsheng Yang, Shulin Li, Kui
Copyright Year	2014
Abstract	NFкB transcription activation leads to malfunction of endothelial cells, which is the main reason for pig xenograft rejection. Overexpression of a dominant negative mutant of porcine NFκB p65 (pp65RHD) could inhibit NFкB activation in endothelial cells. This study presents an advanced tetracycline-regulated system for pp65RHD spatiotemporal expression in porcine iliac endothelial cell line. In this system, an endothelial specific promoter ICAM-2 is used to improve pTet-On and internal ribosome entry site as well as enhanced green fluorescent protein (EGFP) elements are used to facilitate the result observation in pTRE-Tight. Through transfection and drug selection, we obtained 7 single cell clones containing the advanced Tet-On system, in which pp65RHD expression is under tight regulated by doxycycline and can be visualized easily through EGFP. The distribution of induced pp65RHD was verified by immunocytochemical assays test. Then, NFкB activity was tested. Luciferase reporter assays showed that NFкB activity in two clones was influenced by the Dox-induced pp65RHD expression, but other clones weren’t influenced. Therefore, we picked up 2 cell clones from the uninfluenced clones for further investigation by immunocytochemical assays and RT-PCR detection. The final results supported the overexpression of pp65RHD in one clone could successfully inhibit NFкB activity. The success of pp65RHD spatiotemporal expression system is helpful to regulate NFкB activity and conquer cell-mediated immunity and could be used for preparation of transgenic pig, contributing to xenotransplantation.
Starting Page	1753
Ending Page	1761
Page Count	9
File Format	PDF
ISSN	03014851
Journal	Molecular Biology Reports
Volume Number	41
Issue Number	3
e-ISSN	15734978
Language	English
Publisher	Springer Netherlands
Publisher Date	2014-01-18
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Porcine p65RHD PIEC Tet-On advanced system Spatiotemporal expression NFкB activity Animal Biochemistry Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

A retro-lentiviral system for doxycycline-inducible gene expression and gene knockdown in cells with limited proliferative capacity

Expression and SNP association analysis of porcine FBXL4 gene

Several transcription factors regulate COX-2 gene expression in pancreatic β-cells

Molecular characterization and expression patterns of Lbx1 in porcine skeletal muscle

Porcine UCHL1: genomic organization, chromosome localization and expression analysis

Partial molecular characterization, expression pattern, polymorphism and association analysis of porcine SKP2 gene

Molecular characterization and association analysis of porcine interferon regulatory factor 1 gene

Molecular cloning and expression profile analysis of porcine TCAP gene

Molecular cloning, expression patterns and subcellular localization of porcine TMCO1 gene

Spatiotemporal control of porcine p65RHD expression by advanced Tet-On system in PIEC cells helps regulate NFкB activity

Similar Documents

A retro-lentiviral system for doxycycline-inducible gene expression and gene knockdown in cells with limited proliferative capacity

Expression and SNP association analysis of porcine FBXL4 gene

Several transcription factors regulate COX-2 gene expression in pancreatic β-cells

Molecular characterization and expression patterns of Lbx1 in porcine skeletal muscle

Porcine UCHL1: genomic organization, chromosome localization and expression analysis

Partial molecular characterization, expression pattern, polymorphism and association analysis of porcine SKP2 gene

Molecular characterization and association analysis of porcine interferon regulatory factor 1 gene

Molecular cloning and expression profile analysis of porcine TCAP gene

Molecular cloning, expression patterns and subcellular localization of porcine TMCO1 gene

Spatiotemporal control of porcine p65RHD expression by advanced Tet-On system in PIEC cells helps regulate NFкB activity