Method and device for screening sgRNA targets in CRISPR-Cas9 system

Abstract

The invention relates to a CRISPR-Cas9 system sgRNA action target screening method. The method comprises the steps of (1) obtaining segments with 5'-Nx-NGG-3' sequences (x is an integer ranging from 19 to 22 and N represents A / T / C / G) in genomes by utilizing whole genome sequences and gene annotation information of published species to serve as candidate targets of CRISPR-Cas9 system sgRNA; (2) fragmenting the genomes into fragments of 22-25bp and screening sequences ending with NGG and non-repeated on the genomes; and (3) comparing the sequences of the candidate targets in the step (1) with the screened sequences in the step (2), and performing screening and sorting on corresponding optimal sequences according to mismatch information and a selection formula to obtain an optimal genome sgRNA action target set. The invention furthermore provides an apparatus used for realizing the screening method. The method is suitable for all species with known genomes and gene annotation information, and a universal sgRNA sequence set of whole genome level of the species is quickly and efficiently obtained to build a gene knockout mutant library or a gene knockout animal model.

Description

technical field

[0001] The invention relates to the fields of bioinformatics, proteomics, transcriptomics and genetic engineering, in particular to a screening method and device for sgRNA targets of the CRISPR-Cas9 system. Background technique

[0002] With the development of DNA sequencing technology, the genome sequence information of many model organisms has been published, and then researchers turned their research focus to the mining of gene function information. Gene knockout animal models have always been an important tool for studying gene function and finding suitable drug targets in living animals. However, the traditional gene knockout method needs to go through a series of steps such as complex targeting vector construction, embryonic stem cell (ES cell) screening, and chimera breeding. It takes a long time, and the success rate is also affected by many factors. Even for laboratories with relatively mature technology, it generally takes a long time to construct...

Images