BioSkryb, Inc., a developer of genomic amplification technologies, today reported the release of foundational data before publication on the preprint server, bioRxiv, demonstrating the superior ability of primary template-directed amplification (PTA) to accurately and uniformly amplify the genomes of single cells. Whole genome amplification (WGA) is required for low-input DNA and single-cell analysis, but current WGA technologies only cover a fraction of the genome and have a limited capacity to detect genetic variation in each cell. The data show that PTA can overcome these limitations and provide high-quality, scalable analysis of single-cell genomic heterogeneity to enable new insights into human disease at the cellular level and empower high resolution diagnostics for cancer, gene-editing, and other applications.
“We developed PTA to accurately and efficiently decipher genetic diversity at the cellular level so that we can better detect, understand, and treat disease,” said Charles Gawad, M.D., Ph.D., inventor of the PTA technology, cofounder of BioSkryb, and Associate Professor at Stanford University. “These data show that PTA amplifies the genomes of single cells with high coverage breadth and uniformity in a reproducible manner to significantly improve variant calling of all types. The ability of PTA to enable highly accurate genetic information at single-cell resolution could uncover new drug targets for cancer and neurological diseases, as well as identify CRISPR off-target activity to monitor the safety and fidelity of gene editing.”
Single-cell approaches are imperative to tackling difficult biological questions involving genomic heterogeneity, but current WGA methods for single-cell or low-input DNA analysis suffer from significant biases, experimental artifacts, and poor reproducibility. The novel PTA technology accurately calls genomic variants within tissues, one cell at a time, and reproducibly captures more than 95 percent of the genomes of those cells in a more controlled and accurate manner than existing approaches. These data demonstrate that PTA can be reliably used to call copy number variation in primary cancer cells and genome-wide variant detection in gene-edited cells, establishing the feasibility for performing single-cell evaluations of biopsies from cancer and gene-edited tissues.
“Chuck and I cofounded BioSkryb to ensure researchers had access to the most advanced DNA analysis technologies, so it’s especially exciting to see the superior performance of PTA over current methods and to be able to make it commercially available through our ResolveDNA kits,” said Jay West, Ph.D., CEO and cofounder of BioSkryb. “Existing whole genome amplification methods uncover a fraction of the genomic information, but now with ResolveDNA, researchers can explore new places in the genome and be confident in the quality of the sequencing data at those sites.”
ResolveDNA kits are now commercially available and contain all the enzymes and reagents needed for versatile and scalable whole genome amplification from single cells or ultra-low DNA inputs. The kits employ a unique set of nucleotides to prevent recopying of the amplification products, resulting in less errors and higher, more uniform coverage of the genome. The PTA technology also enables attaching cell barcodes to the resultant products for pooling and downstream analysis.
The research paper entitled, “Accurate Genomic Variant Detection in Single Cells with Primary Template-Directed Amplification,” is now available on bioRxiv. The PTA technology was licensed to BioSkryb from St. Jude Children’s Research Hospital, where it was initially developed by Dr. Gawad.
BioSkryb is a venture-backed developer of genomic amplification technologies based in Durham, North Carolina. Its proprietary Primary Template-directed Amplification (PTA) system delivers reproducible, high coverage breadth, and uniformity for various low or ultra-low input sequencing applications. BioSkryb’s products support sequencing library generation produced from limiting samples that have undergone whole genome amplification, resulting in the highest quality genetic analyses available today. For more information, please visit: https://www.bioskryb.com/.