Spatial transcriptome sequencing and decoding method based on transient coding

Abstract

The invention discloses a spatial transcriptome sequencing and decoding method based on transient coding. The method comprises the following steps: designing spatial information coding microspheres based on oligonucleotide chain coding; designing and using a transient coding primer; treating tissue slices and capturing messenger RNA; and building a library, sequencing, analyzing and positioning. According to the method, high-resolution spatial transcriptome sequencing can be obtained, and the sub-cellular resolution (5 [mu]m) can be achieved under the limiting condition, so that the gene expression condition on a specific site in a tissue slice can be accurately obtained. The method is low in cost, high-cost optical spatial position information decoding is avoided, and spatial transcriptome sequencing can be carried out with the cost close to that of single cell transcriptome sequencing.

Description

technical field [0001] The present invention relates to a decoding method for two-dimensional spatial position information of spatial transcriptome sequencing, in particular to a spatial transcriptome sequencing method for obtaining tissue slice transcriptome information and two-dimensional spatial position information at one time through high-throughput sequencing. Background technique [0002] The view that "cells are the basic unit of life" has long been the consensus of many researchers. For multicellular organisms, although the function of a single cell is coordinated and controlled by the whole, each cell is an independent, self-controlled, highly ordered metabolic system, and the metabolic activities of the entire body are carried out with cells as units. It has been found that for individual individuals of most organisms except single-celled organisms, different cell subtypes in various tissues in the body contain approximately the same set of genetic information, th...

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Problems solved by technology

[0003] However, most of the traditional sequencing samples use population cells, which will lead to the final transcriptome information obtained as the average reads of the entire tissue sample, masking the heterogeneity between cells and the discovery of new cell types

Moreover, for the same type of cells, there is also significant microscopic heterogeneity among different individuals, which makes it difficult to reflect the rules of life interaction at the level of a single cell

At the same time, when the tissue or organ under study is manipulated, the first step is to enzymatically hydrolyze it into a cell suspension, which will result in the loss of positional information between cells, making it impossible to obtain the final transcriptome of a single cell. Linking with its location in the tissue is not conducive to researchers studying and understanding the interaction between cell subtypes in the tissue

Most of the existing spatial transcriptome sequencing methods rely on the use of fluorescent probes for multiple rounds of in situ hybridization or in situ sequencing, often requiring a set of expensive optical platforms and a long decoding time

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