History of DNASU

In 2000, Dr. Joshua LaBaer's laboratory at the Harvard Medical School Institute of Proteomics initiated a major effort to create a sequence-verified collection of full-length cDNAs representing all coding regions for Homo sapiens in a vector system that is protein expression-ready for use in high throughput (HT) applications. These clones were shared with researchers around the world through an online ordering system termed Plasmid.

These collections exploit recombinational cloning technology. Conceptually, the cDNAs are captured in a master vector, which positions them to be transferred to virtually any protein expression vector for any use in a single high yield step (in one reaction). By allowing HT automation, a library of genes can be transferred easily and faithfully to a common experimental plasmid for use in proteome scale studies. Using this method, the laboratory has created tens of thousands of plasmids and collections of plasmids, including whole genome collections for several organisms, that can be used in a variety of experimental applications.

In 2006, Plasmid became the Protein Structure Initiative-Materials Repository (later renamed PSI:Biology-Materials Repository) with the mission of providing centralized storage, maintenance and distribution of plasmid clones produced by PSI researchers. PSI was a National Institute of General Medical Sciences (NIGMS) program directed at high throughput structural biology and was composed of structural genomics centers that produced over 100,000 plasmids to be deposited and distributed by the PSI-MR. The shared central goal was to collect these plasmids, fully sequence validate them and distribute them worldwide.

During June 2009, Dr. Joshua LaBaer moved to the Biodesign Institute at Arizona State University. The repository was duplicated and began distribution from ASU in October of 2009 as DNASU.

The next phase of the PSI, termed "PSI:Biology," focused on solving protein structures and applying high-throughput structural biology to important biological problems, which included encouraging partnerships between structural biologists and investigators from the biological, biochemical, and/or molecular communities. This phase of the PSI completed its activity in 2015.

In February 2014, over 82,000 PSI plasmids and their associated data were available upon request through DNASU. In the absence of PSI:Biology-MR, DNASU continued its mission of sharing these plasmids with researchers.

Between 2015-2019, DNASU worked on improvements to the human and viral clone collections with additional support from NIGMS. We reanalyzed our previous collection, Human Orfeome 8.1, in order to compare it with the most current human reference sequence available. For this reanalysis, we identified the open coding regions and binned them into families with >90% homology by sequence alignment. To pick one representative isoform for each gene, we identified the longest isoform and selected the sequence that contained at least 90% of the coding region of this longest isoform. We termed this version of human sequences as the 90/90 collection (90% homology, at least 90% of the longest isoform).

Using these criteria, an analysis of the Human Orfeome 8.1 collection, that listed 12,873 clones, represented only 8,531 of the 20,005 predicted coding regions in RefSeq v92. Thus, this analysis showed that many representative clones were missing. We focused on establishing the missing human clones so that we could offer a more extensive and complete 90/90 collection.

In addition to human genes, we have been creating membrane protein collections from bacteria known to inhabit the gut or respiratory microbiome. Along with the bacterial membrane clones, we have created clones representing complete proteomes of many viruses that are associated with human disease.

During 2020, the COVID pandemic had significant impacts on our operations. Our informatics support team was tasked with establishing the informatics system that supported the ASU Biodesign Clinical Testing Lab, to respond to COVID-19. The wet lab supervisor of DNASU further supported the establishment of the clinical testing lab full time for a period of nine months, leaving DNASU wet lab operations running with two technicians for most of the year. Despite these personnel challenges, DNASU maintained a steady distribution schedule throughout the year, with only minor delays in order deliveries.

While operations seemed to get back to normal for 2021, DNASU experienced multiple challenges but was able to assemble a version 3 of the 90/90 Human ORFeome collection. With the current 90/90 Human ORFeome V3 release, we now have 17,731 clones representing unique genes in the human collection.

We received additional support from NIGMS and with an administrative supplement we will be adding a new automated storage to assist with distribution and housing of our clone collections.

Currently for 2022

The repository has expanded and presently houses and distributes over 320,000 plasmids with inserts from over 1,350 organisms in over 750 vector backbones. These plasmid constructs continue to be deposited from both internal and external depositing consortia and laboratories.

We ship hundreds of clones each week.

We are in the process of transitioning from our current decade old freezer storage unit, Brooks BioStore, into a new Arktic -80 degrees automated storage system. This new system allows for the easy incorporation of additional units in the future as DNASU continues to grow.

We are making much needed updates to our website in both its functionality as well as content. The intent is to create a more up-to-date, user-friendly portal for our clients to place orders, have questions answered, and access supporting data and publications.

We still receive generous support from Dr. Joshua LaBaer who is currently the Executive Director and Professor for the Biodesign Institute, Center Director for Biodesign Virginia G. Piper Center for Personalized Diagnostics, and Interim Center Director for ASU-Banner Neurodegenerative Disease Research Center.

Looking Forward

We expect to announce new versions of current collections.

We are working toward creating a Next Generation Sequencing (NGS) pipeline using tagmentation to sequence verify the entire collection of existing clones in our inventory to maintain integrity and increase reliability of the collections. This undertaking will include not only the gene insert but the vector backbone as well.

We are working to increase our visibility in the scientific community in order to create new external relationships. Through these new relationships, we expect to increase our inventory offerings by the thousands of new clones each year.

Last updated: April 2022