Introduction of a rabbit β-globin gene into the mouse germ line

Costantini, F; Lacy, Elizabeth

doi:10.1038/294092a0

Cited by 432 publications

(143 citation statements)

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“…Although initial reports from a number of laboratories described low levels of globin expression in transgenic mice comparable to that in in vitro experiments (12,25,42,47), one study (43) (14). High-level transcription of viral RNA requires the viral enhancer, and therefore, as expected, near normal levels of viral RNA are seen after differentiation of MEL cells infected with pSVX3(RO) but not with pSVX,Ben-.…”

Section: Resultsmentioning

confidence: 99%

Regulated expression of a complete human beta-globin gene encoded by a transmissible retrovirus vector.

Cone¹,

Weber-Benarous²,

Baorto³

et al. 1987

Mol. Cell. Biol.

150

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We introduced a human 0-globin gene into murine erythroleukemia (MEL) MATERIALS AND METHODSPlasmid constructions. A detailed account of the construction of pSVX has been presented previously (6). pSVXenwas constructed by first digesting a single long terminal repeat (LTR)-containing plasmid completely with XbaI and partially with PvuIl and then religating the appropriate fragment after addition of SalI linkers. A fragment containing the deleted LTR was then introduced in place of the 3' LTR of pSVX. All P-globin vectors were constructed by inserting a BclI-linkered 3.0-kb human 3-globin fragment, from the HpaI site to the PstI site, into the BamHI site of pSVX or pSVXen-. The human ,-globin gene and the SP6 plasmids used for synthesis of probes complementary to mouse and human f-globin mRNAs were kindly provided by

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Section: Resultsmentioning

confidence: 99%

Regulated expression of a complete human beta-globin gene encoded by a transmissible retrovirus vector.

Cone¹,

Weber-Benarous²,

Baorto³

et al. 1987

Mol. Cell. Biol.

150

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“…In transgenic mice obtained by others with several different genes, expression was low or absent [16][17][18] . Consistently successful expression was only obtained with fusion genes of the metal-lothionein (MT) promoter and either the herpes simplex thymidine kinase (tk) gene or the rat growth hormone gene [4][5][6] .…”

Section: Discussionmentioning

confidence: 99%

Expression of a microinjected immunoglobulin gene in the spleen of transgenic mice

Brinster

Ritchie

Hammer

et al. 1983

Nature

193

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Transgenic mice were produced by microinjection of a rearranged, functional immunoglobulin κ gene into fertilized mouse eggs and implantation of the microinjected embryos into foster mothers. Mice that integrated the injected gene were mated and the DNA, RNA and serum κ chains of their offspring were analysed. The data from offspring of three different transgenic mice indicate that the microinjected gene is expressed in the spleen, but not the liver of mice which inherited the injected gene.Immunoglobulin variable (V) and constant (C) region genes must be rearranged into close proximity before expression into a functional immunoglobulin protein can occur 1,2 . The functional rearrangement process takes place only in the B-lymphocyte lineage. To investigate the control of immuno-globulin gene expression we have produced transgenic mice by microinjection 3 of an immunoglobulin gene into fertilized mouse eggs. Microinjection of a gene into an embryo introduces that gene into every cell of the resultant mouse and often leads to efficient expression of the injected gene [4][5][6] . The presence of the injected gene in germ cells allows offspring of transgenic mice to inherit the injected gene. Compared with cell transfection experiments, this approach has the advantages of avoiding potential problems due to the transformed phenotype of cultured cells and allowing direct comparison of expression in different tissues that have the injected gene at the same location in the genome.In our experiments the injected gene is the functional κ gene of the myeloma . Its variable region, V κ M.21, is rearranged next to the joining gene segment J κ 2 (ref. 8) and is contained in the plasmid pB1-14 (Fig. 1). The V κ M.21 portion of the microinjected gene provides a means to distinguish the expression of the introduced gene from that of the endogenous κ genes, both on the RNA and protein levels. The V κ M.21 gene is a member of the V κ 15 family of V κ genes 9,10 . This gene family has approximately ten members which are sufficiently homologous to cross-hybridize using a V κ M.21 probe 11 . V κ genes of this family are probably rearranged and expressed in some B lymphocytes in normal mice, and transcripts from endogenous genes might be expected to confound the detection of transcripts from the injected gene. However, transcription of homologous endogenous V κ 15 were expressed in a tissue at a normal rate in most cells or at a high rate in a subpopulation of cells. The MOPC-21 κ protein is also sufficiently distinct in molecular weight 12 and isoelectric point (see below) that it can be distinguished from other κ chains in mouse serum, provided it is produced and secreted in some quantity. Analysis of transgenic mice containing the pB1-14 gene shows that expression of the microinjected κ gene is apparent on both the RNA and protein levels. HHS Public Access Production of transgenic miceDNA for injection was prepared as described in Fig. 1. Eggs from (C57BL/6 × SJL)F 1 mice were fertilized with F 1 sperm. The male pronuclei o...

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“…For standard expression constructs, however, the classical pronuclear injection approach described here continues to be the major technique used since it was established in the early 1980s (see refs. [3][4][5][6][7][8] as it is a straightforward method to consistently obtain mice with a single integration site. Targeted transgenesis either knocks out or modifies/replaces genes in situ.…”

Section: Introductionmentioning

confidence: 99%