Patent ID: 11898270
Assignee: HUAZHONG AGRICULTURAL UNIVERSITY
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A porcine whole genome-specific sgRNA library, wherein the library includes:
(i) N kinds of vectors expressing porcine-specific sgRNA, wherein N is a positive integer ≥20,000, and the porcine-specific sgRNA is a target gene for the porcine whole genome and the target gene is selected from the group consisting of: (a) a protein encoding gene, (b) a lincRNA, (c) a miRNA and (d) a combination of (a), (b) and (c);
in addition, the porcine-specific sgRNA has the following structural characteristics:
(1) the target gene locus targeted by the sgRNA contains PAM as NGG (wherein N is any base of A, T, C, G);
(2) the length of sgRNA is 19 or 20 nt;
(3) the GC content of sgRNA is 40-60%;
(4) a whole-genome off-target evaluation of sgRNA selects off-target sites that do not contain 1 and 2 base mismatches, compared with other sgRNAs, sgRNAs with a smallest number of off-targets are selected;
(5) for a porcine-specific sgRNA that targets a protein-coding gene, the binding position thereof is located in a region 500 bp downstream of the start codon ATG of the coding gene; for a porcine-specific sgRNA that targets a lincRNA gene, the binding position thereof is located in a 500 bp region downstream of the lincRNA transcription start point; for a porcine-specific sgRNA that targets a miRNA gene, the binding position thereof is a miRNA precursor sequence or a mature sequence; and (ii) optionally, M kinds of negative control vectors, which are the vectors expressing unrelated sgRNA, wherein M is a positive integer.