Meet the new standard in transfection

The new Viromer® PLASMID – more powerful, more potent. The next generation in plasmid transfection.

Viromer PLASMID outperforms major competitors.

Viromer® PLASMID improves pDNA transfection.

Hek293

C2C12

HepG2

A549

A431

Neuro2A

Comparative transfection efficacy of Viromer® PLASMID versus main competitors. Transfection efficacy was measured 24 hours after transfection in various cell lines. Transfections have been done under standard conditions to the manufacturers‘ recommendations for each reagent. (luciferase encoding plasmid (3.5kb) / 100 ng DNA per 96-well / n=4). Viromer® PLASMID shows a strong improvement in plasmid transfection in various cell lines compared to main competitors.

cell line
Fold protein expression improvement of Viromer® PLASMID vs Viromer® RED

HepG2

14

A431

9

C2C12

21

A549

2

Neuro2A

5

Hek293

2

Viromer® RED is a 5-year reference reagent with validated efficiency for plasmid transfection among a very broad range of cells.

Ordering Information

product #
reactions in 24-well

VpD-01LB-00

100

VpD-01LB-01

900

VpD-01LB-03

3 x 900

Unlock transfection limits with mRNA

transient protein expression   |   reprogramming   |   CRISPR / Cas applications   |   immunotherapy assays

Transfection of plasmid DNA is the most common method to overexpress proteins in cells grown in culture. When it fails, not uptake / delivery but rather it‘s processing once into the cell is the hurdle. Reaching and penetrating the nucleus, transcription and release of mRNA to the cytosol are all critical steps.

Transfecting cells with mRNA sequences rather than plasmid DNA constructs gives a great chance to significantly increase transient protein expression levels in a majority of cell types, and offers a unique alternative for challenging cells.

mRNA is a very productive template!

mRNA instructs cells to produce proteins directly. With an optimized delivery it is possible to reach very high expression levels of any intracellular, transmembrane or secreted protein.

mRNA is faster!

No need for transcription! Translation occurs through a promoter-independent process and the desired protein is detectable as early as 2 – 6 h posttransfection.

mRNA is safer!

There is no risk of genomic integration and transient expression avoids toxicity related to protein accumulation.

Superior and rapid expression with 2 times less mRNA than pDNA!

Intensity of fluorescence measured within 52h after transfection of plasmid encoding fluorescent fused protein targeting the outer mitochondrial membrane (pCLOVERTOMM20-N-10) and mRNA-GFP-PDHA1 signal peptide in A549 cells. (pDNA 5kb, mRNA 1,100nt, 96-well)

mRNA transfection enables much stronger protein expression in immune cells

Comparative transfection efficiency of Viromer® RED used for plasmid DNA (light red) or mRNA (dark red) delivery in diverse immune cell lines (maximal percentage of positive cells reported by users).

Comparative GFP expression after plasmid or mRNA transfection using Viromer® RED

primary vascular SMCs

plasmid DNA

mRNA

Data from M. Grossi, Prag, CZ

primary aortic SMCs

plasmid DNA

mRNA

Data from L. Ungethum,
Maastricht University, NL

SH-SY5Y

plasmid DNA

mRNA

Data generated by H. Cynis, Halle, DE

C2C12

plasmid DNA

mRNA

Data generated by H. Cynis, Halle, DE

HUVEC

plasmid DNA

mRNA

Data generated by H. Cynis, Halle, DE

Dendritic cells (hMDDCs)

plasmid DNA

mRNA

Data from F. Gueugnon,
Vaxeal Research, CEA-Saclay, FR

The new Viromer® mRNA outperforms major competitors

Hek293

HepG2

HeLa

MDCK

H358

Neuro2A

A premium high-performance transfection reagent.

Transfection efficacy of Viromer® mRNA compared to competitors.

Luciferase expression was measured 24 hours after transfection in various
cell lines following standard manufacturers‘ protocols.
Luciferase encoding mRNA (1,000nt), 100 ng mRNA per 96-well (n=4)

 

Note: Viromer® RED is a 5-year reference reagent with validated efficiency for both plasmid and mRNA transfection among a very broad range of cells.

Ordering Information

product #
reactions in 24-well

VmR-01LB-00

100

VmR-01LB-01

900

VmR-01LB-03

3 x 900

Superior Transfection for Genome Editing

Maximize genome-editing, transfect RNP with Viromer® CRISPR

» Powerful transfection mediated by high-tech polymer nanoparticles
» Designed for direct delivery of Cas proteins complexed with guide RNAs
» Superior to plasmid or viral approaches, less off-target effects relative to nuclease kinetics
» Low impact on cell viability and physiology
» Comfort of an adjustable and scalable chemical reagent, easy gRNA screening, usable for HTS
» Higher reliability and efficiency of final editing compared to lipofection or electroporation

Delivery of pre-formed RNP complex is safer and faster than plasmid-based transfection:

CRISPR components directly active upon transfection
» No need to use the cell machinery
» Skips the assemblage step in the cytosol

Better control of the Cas9 activity
» Adjustable amounts of delivered Cas9 and gRNA
» R apid protein clearance
» Transient action limiting off-target cleavage
» No integration of Cas9 gene into the cell genome

Safe delivery and 50% of targeted editing in C2C12

Detection of crRNA:tracrRNA-ATTOTM550 (IDT) 24h after RNP delivery

T7E1 assay 72h after RNP delivery ND: no T7 digestion, D: with T7

• Selection of gRNA among 3 synthetic constructs purchased from IDT
• Max. 50% of desired editing achieved with WT Cas9 purified in the lab, while previous assays using plasmid approach showed only 10% of efficiency.

Data courtesy of Dr. Laurence Neff, CMU – University of Geneva, Switzerland

>60% reduction of RFP expression in stable HEK293T

• KO of RFP expression 48h after RNP delivery (final conc. 12.5nM)
• Use of the assay for Viromer library screening
• Parallel comparison with LipofectamineTM CRISPRMAXTM showed 10-20% of efficiency.

Data courtesy of OriGene/US (incl. Cas9 and gRNA sources)

Excess of gRNA leads to >80% of editing in HUVECs

• Sanger sequencing 24h after RNP delivery shows 80-90% of editing with expected +1 insertion as predominant InDel
• Max. effect with 6.25 nM RNP and excess of gRNA (ratio to Cas9 1:2.5)

Cas9 from IDT, gRNA targeting SMAD3 gene from Synthego. External data, contributor not disclosed

Optimization… up to 70% of efficiency in A549

17-72% of cleavage efficiency as results of T7E1 assays 48h after RNP delivery using a gRNA targeting HPRT1 and 2 different Cas9

Data generated by Lipocalyx, Halle, Germany

Ordering Information

product #
reactions in 24-well

VCr-01LB-00

100

VCr-01LB-01

500

VCr-01LB-03

3 x 500