Hong Jessica M, Gibbons Michael, Bashir Ali, Wu Diana, Shao Shirley, Cutts Zachary, Chavarha Mariya, Chen Ye, Schiff Lauren, Foster Mikelle, Church Victoria A, Ching Llyke, Ahadi Sara, Hieu-Thao Le Anna, Tran Alexander, Dimon Michelle, Coram Marc, Williams Brian, Jess Phillip, Berndl Marc, Pawlosky Annalisa
Google, LLC, Mountain View, CA 94043, USA.
iScience. 2021 Dec 11;25(1):103586. doi: 10.1016/j.isci.2021.103586. eCollection 2022 Jan 21.
We demonstrate early progress toward constructing a high-throughput, single-molecule protein sequencing technology utilizing barcoded DNA aptamers (binders) to recognize terminal amino acids of peptides (targets) tethered on a next-generation sequencing chip. DNA binders deposit unique, amino acid-identifying barcodes on the chip. The end goal is that, over multiple binding cycles, a sequential chain of DNA barcodes will identify the amino acid sequence of a peptide. Toward this, we demonstrate successful target identification with two sets of target-binder pairs: DNA-DNA and Peptide-Protein. For DNA-DNA binding, we show assembly and sequencing of DNA barcodes over six consecutive binding cycles. Intriguingly, our computational simulation predicts that a small set of semi-selective DNA binders offers significant coverage of the human proteome. Toward this end, we introduce a binder discovery pipeline that ultimately could merge with the chip assay into a technology called ProtSeq, for future high-throughput, single-molecule protein sequencing.
我们展示了在构建一种高通量单分子蛋白质测序技术方面取得的早期进展，该技术利用带条形码的DNA适配体（结合物）来识别连接在下一代测序芯片上的肽（靶标）的末端氨基酸。DNA结合物在芯片上沉积独特的、用于识别氨基酸的条形码。最终目标是，在多个结合循环中，一串连续的DNA条形码将识别出肽的氨基酸序列。为此，我们展示了使用两组靶标-结合物对（DNA-DNA和肽-蛋白质）成功进行靶标识别的过程。对于DNA-DNA结合，我们展示了在连续六个结合循环中DNA条形码的组装和测序。有趣的是，我们的计算模拟预测，一小部分半选择性DNA结合物能够显著覆盖人类蛋白质组。为此，我们引入了一种结合物发现流程，该流程最终可能与芯片检测合并为一种名为ProtSeq的技术，用于未来的高通量单分子蛋白质测序。
