Three retroviral constructs containing a full-length human alpha-L-iduronidase (IDUA) cDNA were made. The first, pLIdSN, is designed so that expression of the IDUA cDNA is from the 5' viral long terminal repeat (LTR). The second, pLNCId, is designed to express the IDUA cDNA from the cytomegalovirus (CMV) immediate early promoter, while in the third, pLNTId, the CMV promoter is replaced by a promoter fragment of the mouse CD45 (T200) gene. All vectors transduce resistance to G418 (neomycin). High-titer virus-producing cell lines for these constructs were made by infection of the amphotropic packaging cell line PA317 after transient expression in, and virus rescue from, the ecotropic packaging cell line psi CRE. The high-titer virus-producing cell lines were assayed for absence of helper virus, synthesis of human IDUA, and for integrity of proviral structure. Suitable lines were used as a source of virus to infect two different mucopolysaccharidosis type I (MPS I) skin fibroblast cultures. All three of the recombinant viruses corrected the enzymatic defect in MPS I fibroblasts. Surprisingly, increasing over-expression of IDUA resulted in reduced phenotypic correction of these cells as assayed by intracellular accumulation of 35S-labeled glycosaminoglycan. This was shown to be due to the induction of a phenotype analogous to mild I-cell disease in cells expressing large amounts of IDUA.