Chapter 10

The Use of Intracytoplasmic Sperm Injection (ICSI) for Gene Transfer in Mice

Raúl Fernández-González, Pablo Bermejo-Álvarez, Miriam Pérez-Crespo, Alberto Miranda,, Ricardo, Laguna, Celia Frutos and Alfonso Gutiérrez-Adán

Abstract

Using the mouse model, intracytoplasmic sperm injection (ICSI)-mediated transgenesis has been shown to be a valuable tool for the production of transgenic animals, an essential instrument for basic and applied research in bioscience. This method of transgenesis consists of the microinjection of spermatozoa preincubated with foreign DNA. ICSI of DNA-loaded sperm cells has been shown to mediate mouse transgenesis at high efficiency, especially when sperm cell damage (by freeze-thaw cycles or exposure to detergents) is induced. The greatest advantage of ICSI-mediated transgenesis is that it allows introduction of very large DNA transgenes (e.g., yeast artificial chromosomes), with relatively high efficiency into the genome of hosts, as compared to pronuclear microinjection. In addition, ICSI-mediated transgenesis is associated with (a) low frequencies of embryo mosaicism, one of the major limitations of pronuclear microinjection for the production of transgenic livestock, and (b) a high frequency of Mendelian germline transmission of transgenic sequences among founder animals. In this chapter we will review some factors that can increase the efficiency of the ICSI-mediated transgenesis and we will describe a new active form of ICSImediated transgenesis employing fresh sperm in conjunction with recombinase or transposase molecules.

Total Pages: 103-111 (9)

Purchase Chapter  Book Details