Aug 29, 2005 · We report that homologous recombination using a promoterless gene trap vector (“targeting trapping”) yields targeting frequencies averaging above 50%, a significant increase compared with current approaches.

Jun 30, 2015 · In this paper, we describe a simple and rapid method to construct exon-trap vectors that allow for efficient targeted gene disruption in mouse and human cells. Furthermore, we develop a novel method to enhance the efficiency of counterselection for non-homologous integration of exon-trap vectors.

Gene trapping is a high-throughput approach that is used to introduce insertional mutations across an organism's genome.

Oct 27, 2021 · Insertion of a gene trap vector disrupts gene function, reports gene expression, and provides a convenient tag for the identification of the insertion site. The trap vector can be delivered to ES cells by electroporation of a plasmid, by retroviral infection, or by transposon-mediated insertion.

Gene trap constructs are typically introduced into totipotent embryonic stem cells by using a retroviral vector. Embryonic stem cells that subsequently test positive for retroviral integration are subsequently transferred to the mouse germline.

Mar 1, 2006 · In this article, the origin and background of gene trapping technology are briefly introduced, and then the principles of typical gene trap vectors, current progress and new efficient gene trapping techniques are discussed in detail. Disadvantage and prospects of various gene trapping technologies are also discussed.

Aug 25, 2000 · For instance Skarnes and collaborators [14] have planned a novel gene trap strategy, named secretory trap, which is based on a new class of vectors (see above) and this subcellular localization prescreening.

In this paper, we outline methods for constructing targeting vectors for efficient genetic modification, with an emphasis on exon-trap vectors and a novel negative selection strategy to enhance gene targeting in human cells. Fig. 1. Gene Targeting via Homologous Recombination Is Inefficient in Human Cells.

Jan 1, 2011 · The international knockout mouse project (IKMC) uses a “knockout first” targeting vector design. The targeting vector includes a splice acceptor designed to substitute the expression of a lacZ cassette for the expression of the endogenous gene 5′ of a floxed exon.

Feb 28, 2018 · Gene trapping is a type of insertional mutagenesis that disrupts gene function by the integration of a vector in the intergeneric sequences.