Patent ID: 8722408
Filing Date: 2014-05-13
Classification: C12N

Abstract:
1. A method of gene trapping, comprising the steps of: introducing a trap vector into mouse embryonic stem cells, the trap vector comprising: 1) a loxP sequence and a lox71 sequence, wherein the loxP sequence comprises in sequential order inverted repeat sequence 1, a spacer sequence, and inverted repeat sequence 2; wherein the loxP71 sequence comprises the nucleotide sequence showin in SEQ ID NO: 13; and wherein the trap vector is suitable for use in mammalian cells, or 2) a loxP sequence and a lox66 sequence, wherein the loxP sequence comprises in sequential order inverted repeat sequence 1, a spacer sequence, and inverted repeat sequence 2, wherein the lox66 comprises the nucleotide sequence shown in SEQ ID NO: 16, and wherein the trap vector is suitable for use in mammalian cells, wherein the trap vector comprises a splice acceptor site and an internal ribosomal entry site; culturing the mouse embryonic stem cells; selecting those cells which exhibit a pattern of single copy integration of the trap vector; and isolating the trapped gene.