Department of Genome Sciences, University of Washington, Seattle, WA, USA.
Allen Institute for Brain Science, Seattle, WA, USA.
Nat Methods. 2021 Aug;18(8):937-944. doi: 10.1038/s41592-021-01187-3. Epub 2021 Jul 5.
Fluorescence in situ hybridization (FISH) allows researchers to visualize the spatial position and quantity of nucleic acids in fixed samples. Recently, considerable progress has been made in developing oligonucleotide (oligo)-based FISH methods that have enabled researchers to study the three-dimensional organization of the genome at super-resolution and visualize the spatial patterns of gene expression for thousands of genes in individual cells. However, there are few existing computational tools to support the bioinformatics workflows necessary to carry out these experiments using oligo FISH probes. Here, we introduce paint server and homology optimization pipeline (PaintSHOP), an interactive platform for the design of oligo FISH experiments. PaintSHOP enables researchers to identify probes for their experimental targets efficiently, to incorporate additional necessary sequences such as primer pairs and to easily generate files documenting library design. PaintSHOP democratizes and standardizes the process of designing complex probe sets for the oligo FISH community.
荧光原位杂交(FISH)使研究人员能够在固定样本中可视化核酸的空间位置和数量。最近,在开发基于寡核苷酸(oligo)的 FISH 方法方面取得了相当大的进展,这些方法使研究人员能够以超高分辨率研究基因组的三维结构,并可视化单个细胞中数千个基因的空间表达模式。然而,现有的计算工具很少能够支持使用 oligo FISH 探针进行这些实验所需的生物信息学工作流程。在这里,我们介绍了 paint server 和同源优化管道(PaintSHOP),这是一个用于设计 oligo FISH 实验的交互式平台。PaintSHOP 使研究人员能够有效地为他们的实验目标识别探针,合并额外的必要序列,如引物对,并轻松生成记录库设计的文件。PaintSHOP 使设计 oligo FISH 社区中复杂探针集的过程民主化和标准化。