Devgan, Vikram and Thomas, Monzy and Ullas, Kolathur S and Rao, Manchanahalli RS and Seshagiri, Polani B (2003) Embryo culture-based generation of enhanced green fluorescent protein-transgenic mice. In: Biochemical and Biophysical Research Communications, 303 (4). pp. 994-1001.
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One of the limitations of transgenesis is low efficiency. In this study, we generated transgenic mice harboring the enhanced green fluorescent protein (EGFP) gene, under the control of chicken-β-actin promoter and cytomegalovirus enhancer, using two approaches and compared their efficiencies. One involved culture of EGFP-injected embryos developing through EGFP-expressing “green” blastocysts, followed by their transfer to uterus. The second was oviductal-transfer of EGFP-injected-eggs. Embryo culture-based-transgenesis (ECBT) produced 100% transgenic mice, unlike the second approach. Moreover, ECBT required reduced number of recipients and markedly increased pregnancy rates. Of the nine founders, seven exhibited ubiquitous EGFP-expression, one (GU1) was a mosaic and the other (G18) was non-expressing. The molecular basis for this was attributed to repeat-induced gene silencing, since the G18 had a high copy number ( 99/genome) of the non-mutated and non-rearranged EGFP-transgene integrated at a single site. Our results show the superiority of ECBT over the conventional oviductal approach for generating transgenic “green” mice.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Elsevier Science|
|Keywords:||Embryo culture based transgenesis;Enhanced green fluorescent protein;Transgenic mouse;Mosaicism; Gene silencing|
|Department/Centre:||Division of Biological Sciences > Molecular Reproduction, Development & Genetics (formed by the merger of DBGL and CRBME)
Division of Biological Sciences > Biochemistry
|Date Deposited:||13 Oct 2007|
|Last Modified:||19 Sep 2010 04:40|
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