The Power of Small

December 14, 2022 / by Patrick Paez, Ph.D.

Whitepaper showcases next-generation plasmid technology

Nanoplasmid^TM. We’ve been discussing this topic quite a bit recently, and now our latest whitepaper brings you an in-depth examination of our novel Next-Generation plasmid technology. The etiology of its name, “nano,” originates from its purposefully designed “small” backbone of around 500 base pairs (bp) as compared to traditional plasmids that often exceed 2000 bp. This reduced backbone size is a direct result of removing prokaryotic and antibiotic-resistance genes used to select transformed bacteria and propagate a vector.

What’s this about, “The Power of Small”?
Given the size reduction, Nanoplasmid reduces transfection toxicities and innate immune genes activated by the presence of prokaryotic genes rampantly expressed by traditional plasmids. Its smaller backbone increases gene expression, reduces transgene silencing, and provides an improved patient safety profile by eliminating the possibility of residual antibiotics contaminating a final plasmid preparation, eliminating the chance of antibiotic-resistance gene transfer, or the propagation of a plasmid in a patient’s commensal bacteria (i.e., the gut microbiome) through R6K strain dependence.

The benefits of this next-generation plasmid are pleiotropic and applicable to a wide array of applications, such as:

• Viral vectors such as AAV and lentivirus
• Non-viral vectors like CRISPR homology-directed repair (HDR) as a donor template
• Transposons as a donor template
• As a template for mRNA production
• Novel episomal scaffold/matrix attachment motifs used to maintain the replication of a vector in dividing cells

Improved Transfection efficiencies
One of the first studies examined in the whitepaper compares a traditional plasmid vector to Nanoplasmid. Results showed an improvement in transfection efficiency, reduced transfection-associated toxicities, and the ability to reduce electroporation voltages in vivo and in vitro when using Nanoplasmid without compromising performance. Transfections constitute a significant step in many applications, and improving their efficiencies can help reduce manufacturing costs and improve final product yields.

Application-based data using Nanoplasmid
In another study, Genentech Inc. used Aldevron’s SpyFiTM CRISPR/Cas9 to HDR knock-in the NY-ESO-1 specific TCR into human T cells. The study compared knock-in efficiencies between these templates:

• Linear double-stranded DNA (dsDNA)
• Traditional pUC plasmid
• Nanoplasmid, our next-generation plasmid vector

The results showed that Nanoplasmid outperformed both the more costly linear dsDNA and the traditional plasmid pUC vector, increasing knock-in efficiencies, improving cell viability after electroporation, and enhancing T cell phenotypes after recovery.

See more published applications using Nanoplasmid
Download our whitepaper today to learn more about the Nanoplasmid vector’s benefits for other applications, including mRNA and DNA vaccines, AAV and lentivirus packaging and production, and more cases of CRISPR and Transposon therapies.

DOWNLOAD NOW

• Want to learn more about this topic? Contact Patrick Paez
• Visit the Nanoplasmid page of our website