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Nucleic Acids Research, 1992, Vol. 20, No. 23 6287-6295
© 1992

MOLECULAR BIOLOGY

A transgenic mouse that expresses a diversity of human sequence heavy and light chain immunoglobulins

Lisa D. Taylor, Condie E. Carmack, Stephen R. Schramm, Roshanak Mashayekh, Kay M. Higgins, Chiung-Chi Kuo, Clive Woodhouse, Robert M. Kay and Nils Lonberg*

GenPharm International 2375 Garcia Avenue, Mountain View, CA 94043, USA

*To whom correspondence should be addressed

Received August 27, 1992. Revised October 26, 1992. Accepted October 26, 1992.

We have generated transgenic mice that express a diverse repertoire of human sequence immunoglobulins. The expression of this repertoire is directed by light and heavy chain minilocus transgenes comprised of human protein coding sequences in an unrearranged, germ-line configuration. In this paper we describe the construction of these miniloci and the composition of the CDR3 repertoire generated by the transgenic mice. The largest transgene discussed is a heavy chain minilocus that includes human µand {gamma}1 coding sequences together with their respective switch regions. It consists of a single 61 kb DNA fragment propagated in a bacterial plasmid vector. Both human heavy chain classes are expressed in animals that carry the transgene. In light chain transgenic animals the unrearranged minilocus sequences recombine to form VJ joints that use all five human Jx, segments, resulting in a diversity of human-like CDR3 regions. Similarly, in heavy chain transgenics the inserted sequences undergo VDJ joining complete with N region addition to generate a human-like VH CDR3 repertoire. All six human JH segments and at least eight of the ten transgene encoded human D segments are expressed. The transgenic animals described in this paper represent a potential source of human sequence antibodies for in vivo therapeutic applications.


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