Devgan, Vikram ; Thomas, Monzy ; Ullas, Kolathur S ; Rao, Manchanahalli R.S ; Seshagiri, Polani B (2003) Embryo culture-based generation of enhanced green fluorescent protein-transgenic mice Biochemical and Biophysical Research Communications, 303 (4). pp. 994-1001. ISSN 0006-291X
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Official URL: http://doi.org/10.1016/s0006-291x(03)00483-2
Related URL: http://dx.doi.org/10.1016/s0006-291x(03)00483-2
Abstract
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-beta-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 (approximately 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: | Article |
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Source: | Copyright of this article belongs to Elsevier Science |
Keywords: | Embryo Culture-Based-Transgenesis; Enhanced Green Fluorescent Protein; Transgenic Mouse; Mosaicism; Gene Silencing. |
ID Code: | 118234 |
Deposited On: | 19 May 2021 09:17 |
Last Modified: | 19 May 2021 09:17 |
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