Overcoming the DNA Bottleneck

Abstract

Genetic Engineering & Biotechnology NewsVol. 42, No. 6 Sponsored ContentFree AccessOvercoming the DNA BottleneckPowering nucleic acid–based therapies into the future with LinearDNA™Published Online:8 Jun 2022https://doi.org/10.1089/gen.42.06.01AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail The global demand for DNA continues to rise at a breakneck pace. This trend began with the introduction of redirected cell and gene therapies and was supercharged by the success of mRNA COVID-19 vaccines. While these transformative therapies come in a variety of forms, they all have a common thread—they rely on DNA.According to a May 2021 report from the American Society of Gene and Cell Therapy, there were approximately 1,745 gene/cell therapies and 600 RNA therapies in development.1 These numbers have continued to grow over the past year. The net result is an unprecedented demand for DNA for research, clinical development, and therapeutic manufacturing.Plasmid DNA—good enough, until nowTo date, nucleic acid–based therapies have used plasmid DNA (pDNA) as their source of DNA. Based on 1970's technology, pDNA was the only viable source of DNA when nucleic acid–based therapy development began. Thus pDNA became the industry standard.pDNA has several limitations and downsides, including the presence of antibiotic resistance genes and origins of replication, inclusion of unwanted DNA sequences, low yields, batch failures and inconsistencies, as well as sequence heterogeneity—the latter two issues being exacerbated by the complex DNA sequences needed for gene and mRNA therapies.Moreover, pDNA manufacturing is complex, and scaling its manufacture is both time consuming and expensive. These facts have resulted in a pDNA supply that has failed to meet current DNA demands.LinearDNA—breaking the DNA bottleneckLinearDNA is an enzymatically produced DNA that is substitutable for pDNA in nucleic acid–based therapy manufacturing. LinearDNA is produced via the polymerase chain reaction (PCR) in a cell-free manufacturing workflow, and unlike plasmids, contains no extraneous DNA sequences. LinearDNA can be produced from an existing pDNA construct or a de novo DNA sequence.LinearDNA is substitutable for pDNA in a wide range of nucleic acid–based therapy applications, including mRNA production, CAR-T and viral vector manufacturing, and DNA vaccines. LinearDNA is produced via a rapidly scalable manufacturing platform that can grow to meet the needs of virtually any customer.Gram-scale quantities of LinearDNA can be produced in mere weeks with high sequence fidelity and homogeneity—even with complex DNA sequences such as polyA and inverted terminal repeats (ITRs). Recently, a proof-of-concept study was conducted in which a commercial-scale batch of LinearDNA was produced from a de novo DNA template in 10 days—highlighting LinearDNA's unparalleled speed of production.Each batch of LinearDNA is produced to high purity according to strict quality specifications as shown in the table above.LinearDNA—powering mRNA therapies into the futureLinearDNA can accelerate and simplify mRNA research and production workflows that currently use pDNA. Unlike pDNA, which must be “linearized” via expensive restriction enzymes for in vitro transcription (IVT), LinearDNA is ready for IVT use without further modification. In addition, since LinearDNA is free of pDNA backbone sequences, there are no adventitious DNA sequences to remove.Furthermore, unlike plasmid-based DNA production methods that produce heterogeneous DNA sequences when faced with polyA sequences, large quantities of homogeneous LinearDNA can be produced that include a precise number of polyA nucleotides (ranging from 80 to 120).The combination of these unique advantages, coupled with LinearDNA's rapid production times and high purity, makes LinearDNA an ideal DNA template to continue the rapid growth of mRNA therapies.ConclusionpDNA, once the enabler of nucleic acid–based therapies, is now a problematic bottleneck. LinearDNA can rapidly meet all current and future DNA demands and remove DNA supply constraints on therapy developers and manufacturers.Learn more about LinearDNAadnas.com/lineardnaReference1. American Society of Gene and Cell Therapy. Gene, Cell & RNA Therapy Landscape: Q2 2021. Google ScholarFiguresReferencesRelatedDetails Volume 42Issue 6Jun 2022 Information© 2022 by GEN PublishingTo cite this article:Overcoming the DNA Bottleneck.Genetic Engineering & Biotechnology News.Jun 2022.7-7.http://doi.org/10.1089/gen.42.06.01Published in Volume: 42 Issue 6: June 8, 2022PDF download