Cloning of apoB intrabodies: specific knockdown of apoB in HepG2 cells.
作者： Wei Liao , Randall W. Strube , Ross W. Milne , Si-Yi Chen , Lawrence Chan
DOI: 10.1016/J.BBRC.2008.06.020
关键词: Single-Chain Antibodies 、 Endoplasmic reticulum 、 Biology 、 Intracellular 、 Apolipoprotein B 、 Secretion 、 Molecular biology 、 Intrabody 、 Gene knockdown 、 Antibody
摘要: Apolipoprotein (apo) B is essential for the assembly and secretion of triglyceride-rich lipoproteins made by liver. As sole protein component in LDL, apoB an important determinant atherosclerosis susceptibility a potential pharmaceutical target. Single-chain antibodies (sFvs) are smallest fragment IgG molecule capable maintaining antigen binding specificity parental antibody. In present study, we describe cloning construction two intracellular (intrabodies) to human apoB. We targeted these intrabodies endoplasmic reticulum purpose retaining nascent within ER, thereby preventing its secretion. Expression 1D1 intrabody apoB-secreting hepatoma cell line HepG2 resulted marked reduction This study demonstrates utility specifically block plasma LDL as therapeutic strategy treatment hyperlipidemia.
elsevier.com
sci-hub.se 参考文章(13)
R J Pease, R W Milne, W K Jessup, A Law, P Provost, J C Fruchart, R T Dean, Y L Marcel, J Scott, Use of bacterial expression cloning to localize the epitopes for a series of monoclonal antibodies against apolipoprotein B100. Journal of Biological Chemistry. ,vol. 265, pp. 553- 568 ,(1990) , 10.1016/S0021-9258(19)40266-4
W. A. Marasco, W. A. Haseltine, S. Y. Chen, Design, intracellular expression, and activity of a human anti-human immunodeficiency virus type 1 gp120 single-chain antibody Proceedings of the National Academy of Sciences of the United States of America. ,vol. 90, pp. 7889- 7893 ,(1993) , 10.1073/PNAS.90.16.7889
Munechika Enjoji, Fang Wang, Makoto Nakamuta, Lawrence Chan, Ba-Bie Teng, Hammerhead Ribozyme as a Therapeutic Agent for Hyperlipidemia: Production of Truncated Apolipoprotein B and Hypolipidemic Effects in a Dyslipidemia Murine Model Human Gene Therapy. ,vol. 11, pp. 2415- 2430 ,(2000) , 10.1089/104303400750038516
H. Jamil, D. A. Gordon, D. C. Eustice, C. M. Brooks, J. K. Dickson, Y. Chen, B. Ricci, C. H. Chu, T. W. Harrity, C. P. Ciosek, S. A. Biller, R. E. Gregg, J. R. Wetterau, An inhibitor of the microsomal triglyceride transfer protein inhibits apoB secretion from HepG2 cells Proceedings of the National Academy of Sciences of the United States of America. ,vol. 93, pp. 11991- 11995 ,(1996) , 10.1073/PNAS.93.21.11991
Wei Liao, Benny Hung-Junn Chang, Michael Mancini, Lawrence Chan, Ubiquitin-dependent and -independent proteasomal degradation of apoB associated with endoplasmic reticulum and Golgi apparatus, respectively, in HepG2 cells. Journal of Cellular Biochemistry. ,vol. 89, pp. 1019- 1029 ,(2003) , 10.1002/JCB.10538
R. Orlandi, D. H. Gussow, P. T. Jones, G. Winter, Cloning immunoglobulin variable domains for expression by the polymerase chain reaction. Proceedings of the National Academy of Sciences of the United States of America. ,vol. 86, pp. 3833- 3837 ,(1989) , 10.1073/PNAS.86.10.3833
Si-Yi Chen, Jessamyn Bagley, Wayne A. Marasco, Intracellular Antibodies as a New Class of Therapeutic Molecules for Gene Therapy Human Gene Therapy. ,vol. 5, pp. 595- 601 ,(1994) , 10.1089/HUM.1994.5.5-595
Wei Liao, Guang Ning, Knockdown of apolipoprotein B, an atherogenic apolipoprotein, in HepG2 cells by lentivirus-mediated siRNA. Biochemical and Biophysical Research Communications. ,vol. 344, pp. 478- 483 ,(2006) , 10.1016/J.BBRC.2006.03.164
Isaac J. Rondon, and Wayne A. Marasco, INTRACELLULAR ANTIBODIES (INTRABODIES) FOR GENE THERAPY OF INFECTIOUS DISEASES Annual Review of Microbiology. ,vol. 51, pp. 257- 283 ,(1997) , 10.1146/ANNUREV.MICRO.51.1.257
Jürgen Soutschek, Akin Akinc, Birgit Bramlage, Klaus Charisse, Rainer Constien, Mary Donoghue, Sayda Elbashir, Anke Geick, Philipp Hadwiger, Jens Harborth, Matthias John, Venkitasamy Kesavan, Gary Lavine, Rajendra K. Pandey, Timothy Racie, Kallanthottathil G. Rajeev, Ingo Röhl, Ivanka Toudjarska, Gang Wang, Silvio Wuschko, David Bumcrot, Victor Koteliansky, Stefan Limmer, Muthiah Manoharan, Hans-Peter Vornlocher, Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs Nature. ,vol. 432, pp. 173- 178 ,(2004) , 10.1038/NATURE03121