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Journal Club

The Mirus Bio Journal Club highlights recent publications and technologies relevant to transfection and gene transfer applications. Its purpose is to increase awareness of innovative research fueled by nucleic acid delivery.

Case Study // JUN 8 2015

Setting the Foundation for an Ebola Vaccine

Generation of a cell line that is biologically contained for Ebola virus research "The VeroVP30 cell line was established by cotransfecting Vero cells with pCAG-VP30 (for the expression of VP30) and pPur, a protein expression plasmid for the puromycin resistance gene (Clontech), using the transfection reagent TransIT®-LT1 (Mirus)..... To artificially generate EBOV, we transfected 5 x 105 293T cells with 1.0 ug of pTM-EbolaΔVP30, 2.0 ug of pCAG-L, 1.0 ug of pCAG-NP, 0.5 ug of pCAG-VP35, 0.5 ug of pCAG-VP30 and 1.0 ug of pCAG-T7 pol using TransIT®-LT1 (Mirus) in BSL-4 containment" - Halfmann, et al.1

Case Study // NOV 10 2014

Mesenchymal stem cells (MSCs) transfected with TRAIL-bearing vectors provide an antitumor therapy

Pancreatic cancer continues to have a very poor prognosis and treatments are elusive due to the lack of tumor specific therapies; however, the tumor-homing characteristic of MSCs provides an attractive vehicle to deliver therapeutics. The current study shows this proof of concept through expression of the anti-cancer agent TRAIL from MSCs transfected using TransIT®-2020.

Case Study // OCT 6 2014

A strategy for more uniform glycoproteins: Keep it simple

Mammalian glycoproteins exist as many differently glycosylated variants. For protein-based drugs such as therapeutic antibodies, the diversity of glycoforms leads to heterogeneity in folding, stability, and immunogenicity. Through cell line engineering it may be possible to make recombinant glycoproteins with smaller and more homogeneous glycans, thereby minimizing intra-batch and batch-to-batch variability for biotherapeutic proteins.

Case Study // SEP 4 2014

High Transient Transfection Levels Translate to Higher Specific Productivity in Suspension CHO cells

Upstream production of biologic drug targets often utilizes transient transfection methods to rapidly generate usable amounts of protein for preclinical studies. This is frequently performed in parallel with the more labor intensive stable cell line development. Transfection methodologies, media formulation, cell line engineering, and vector design are all important factors to enhance yields in upstream development of biologics in suspension CHO cells.

Case Study // JUL 21 2014

Co-transfection of multiple plasmid DNAs encoding the IBV genome paves way for better flu vaccines

Yearly influenza outbreaks impose a huge public health cost as the effectiveness of traditional flu vaccines can vary greatly from one year to the next. Due to the antigenic drift and sometimes antigenic shifts (that can lead to an influenza pandemic), there can be a high degree of mismatch between the vaccine seed stock and the actual seasonal flu virus of a particular season. Obtaining gene sequence information of virus isolates from different geographical origin can lead to smarter design of flu vaccines in the future.

Case Study // JUN 30 2014

Genome editing mediated by efficient transfection of dimeric CRISPR/Fok1 nuclease encoding plasmids

The ability to quickly and easily alter the genome ushers forth an era in which relevant disease models can be created, pathways can be better analyzed and therapeutic possibilities become reality. A variety of genome editing methodologies are available including Zinc Finger Nucleases (ZFNs) and Transcription Activation Effector-like Nucleases (TALENs), but now the use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) has come to the forefront as a much more effective strategy. Transfection, or nucleic acid delivery to the cell, is a necessary requirement of genome editing techniques.