US 12,606,820 B2 · Genome-wide insulator screening system and use

Abstract

The present disclosure discloses a genome-wide insulator screening system. The system comprises an MAI-seq-experiment vector and an MAI-seq-control vector; core gene elements in the MAI-seq-experiment vector are arranged in following order: a weak promoter, a marker target gene, an insertion site for a sequence to be screened, an enhancer and a poly A site; and core gene elements in the MAI-seq-control vector are arranged in following order: a weak promoter, a marker target gene, an enhancer, an insertion site for the sequence to be screened and a poly A site. This genome-wide insulator screening system, primarily composed of these two vectors, exhibits high sensitivity and can be applied to the screening of genomic insulators in any species. Furthermore, it effectively eliminates the influence of silencers, ensuring high screening accuracy. This system provides important technical support for constructing insulator maps and understanding the characteristics and mechanisms of action of insulators.

Claims4

Dependent claims

A genome-wide insulator screening system (MAI-seq), wherein the system comprises an MAI-seq-experiment vector and an MAI-seq-control vector;

core gene elements in the MAI-seq-experiment vector are arranged in the following order:
a weak promoter, a marker target gene, an insertion site for a sequence to be screened, an enhancer and a poly A site;
core gene elements in the MAI-seq-control vector are arranged in the following order:
a weak promoter, a marker target gene, an enhancer, an insertion site for the sequence to be screened and a poly A site;
the insertion site for the sequence to be screened contains homologous arm sequences that are configured to hybridize with complementary sequences attached to the DNA sequence to be screened to enable homologous recombination, and after two ends of a DNA sequence to be screened are added to the homologous arm sequences, homologous recombination occurs with either the MAI-seq-experiment vector or the MAI-seq-control vector;
and a nucleotide sequence of the MAI-seq-experiment vector is as shown in SEQ ID NO:
1, and a nucleotide sequence of the MAI-seq-control vector is as shown in SEQ ID NO:2.

4 more elements

Dependencies are inferred from the claim text.

Genome-wide insulator screening system and use

Life of this patent

Genome-wide insulator screening system and use

Life of this patent

Abstract

Claims4