How can I increase the transduction efficiency with the BacMam 2.0 reagents such as the the CellLight and Premo products?
Try varying particle-to-cell ratio (PPC), incubation volume, temperature and, cell density (if adherent cells are transduced). For adherent cells, we recommend a confluence of about 70%. Following the PPC, adjusting the volume is the next best parameter to change to optimize protein expression. If that doesn't work, you can also use the BacMam Enhancer Kit (Cat. No. B10107).
A Rab11a-enriched subapical membrane compartment regulates a cytoskeleton-dependent transcytotic pathway in secretory epithelial cells of the lacrimal gland.
Authors:Xu S, Edman M, Kothawala MS, Sun G, Chiang L, Mircheff A, Zhu L, Okamoto C, Hamm-Alvarez S,
Journal:J Cell Sci
PubMed ID:21984810
'Despite observations that the lacrimal gland has been identified as the principal source of dimeric immunoglobulin A (dIgA) in tears, the mechanism used by lacrimal gland acinar cells (LGACs) to transcytose dIgA produced by interstitial plasma cells is not well-characterized. This study identifies a transcytotic pathway in LGACs regulated by ... More
Deletion of lipoprotein PG0717 in Porphyromonas gingivalis W83 reduces gingipain activity and alters trafficking in and response by host cells.
Authors:Reyes L, Eiler-McManis E, Rodrigues PH, Chadda AS, Wallet SM, Bélanger M, Barrett AG, Alvarez S, Akin D, Dunn WA, Progulske-Fox A,
Journal:
PubMed ID:24069284
'P. gingivalis (Pg), a causative agent of chronic generalized periodontitis, has been implicated in promoting cardiovascular disease. Expression of lipoprotein gene PG0717 of Pg strain W83 was found to be transiently upregulated during invasion of human coronary artery endothelial cells (HCAEC), suggesting this protein may be involved in virulence. We ... More
Improving drug potency and efficacy by nanocarrier-mediated subcellular targeting.
Authors:Murakami M, Cabral H, Matsumoto Y, Wu S, Kano MR, Yamori T, Nishiyama N, Kataoka K,
Journal:Sci Transl Med
PubMed ID:21209412
'Nanocarrier-mediated drug targeting is an emerging strategy for cancer therapy and is being used, for example, with chemotherapeutic agents for ovarian cancer. Nanocarriers are selectively accumulated in tumors as a result of their enhanced permeability and retention of macromolecules, thereby enhancing the antitumor activity of the nanocarrier-associated drugs. We investigated ... More
Baculovirus-mediated gene transfer into mammalian cells.
Authors:Boyce FM, Bucher NL,
Journal:Proc Natl Acad Sci U S A
PubMed ID:8637876
This paper describes the use of the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a vector for gene delivery into mammalian cells. A modified AcMNPV virus was prepared that carried the Escherichia coli lacZ reporter gene under control of the Rous sarcoma virus promoter and mammalian RNA processing ... More
BacMam technology and its application to drug discovery.
Authors:Ames RS, Kost TA, Condreay JP,
Journal:Expert Opin Drug Discov
PubMed ID:23488908
The recombinant baculovirus/insect cell system was firmly established as a leading method for recombinant protein production when a new potential use for these viruses was revealed in 1995. It was reported that engineered recombinant baculoviruses could deliver functional expression cassettes to mammalian cell types; a system which has come to ... More