Inadvertent transgenesis by conventional ICSI in mice

Background ICSI is a relatively new treatment for human male-related infertility, as well as an efficient method for the production of transgenic animals by injecting into the oocyte sperm previously incubated with foreign DNA. As semen samples collected in human infertility clinics are frequently contaminated with bacteria, one risk associated with the ICSI procedure is the injection of foreign, sperm-associated exogenous DNA into the oocyte, and the generation of transgenic offspring. Methods To analyse this possibility, ICSI was performed in mouse oocytes with frozen-thawed and Percoll-treated fresh sperm samples intentionally contaminated with plasmid EGFP-transformed E. coli bacteria or medium from which these bacteria were washed. Fertilized embryos were cultured in vitro until morula/blastocyst stage, transferred into pseudopregnant females, and at day 14, fetuses and reabsorptions were analysed by PCR for the genomic presence of integrated plasmid and/or bacterial DNA. Results Independently of the sperm pretreatment tested, transgenesis was produced. Conclusions Inadvertent transgenesis by conventional ICSI is a possibility that should not be neglected. Particular precautions, such as full bacteriological semen examinations and effective antibiotic semen pretreatments, should be taken in human infertility clinics, in order to exclude the possibility of accidental transgenesis. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.

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Main Authors: Moreira, P. N., Fernández-González, R., Rizos, D., Ramirez, M., Perez-Crespo, M., Gutiérrez-Adán, A.
Format: journal article biblioteca
Language:eng
Published: 2005
Online Access:http://hdl.handle.net/20.500.12792/2990
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spelling dig-inia-es-20.500.12792-29902020-12-15T09:14:51Z Inadvertent transgenesis by conventional ICSI in mice Moreira, P. N. Fernández-González, R. Rizos, D. Ramirez, M. Perez-Crespo, M. Gutiérrez-Adán, A. Background ICSI is a relatively new treatment for human male-related infertility, as well as an efficient method for the production of transgenic animals by injecting into the oocyte sperm previously incubated with foreign DNA. As semen samples collected in human infertility clinics are frequently contaminated with bacteria, one risk associated with the ICSI procedure is the injection of foreign, sperm-associated exogenous DNA into the oocyte, and the generation of transgenic offspring. Methods To analyse this possibility, ICSI was performed in mouse oocytes with frozen-thawed and Percoll-treated fresh sperm samples intentionally contaminated with plasmid EGFP-transformed E. coli bacteria or medium from which these bacteria were washed. Fertilized embryos were cultured in vitro until morula/blastocyst stage, transferred into pseudopregnant females, and at day 14, fetuses and reabsorptions were analysed by PCR for the genomic presence of integrated plasmid and/or bacterial DNA. Results Independently of the sperm pretreatment tested, transgenesis was produced. Conclusions Inadvertent transgenesis by conventional ICSI is a possibility that should not be neglected. Particular precautions, such as full bacteriological semen examinations and effective antibiotic semen pretreatments, should be taken in human infertility clinics, in order to exclude the possibility of accidental transgenesis. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. 2020-10-22T13:50:12Z 2020-10-22T13:50:12Z 2005 journal article http://hdl.handle.net/20.500.12792/2990 10.1093/humrep/dei237 eng Attribution-NonCommercial-ShareAlike 4.0 International http://creativecommons.org/licenses/by-nc-sa/4.0/ open access
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description Background ICSI is a relatively new treatment for human male-related infertility, as well as an efficient method for the production of transgenic animals by injecting into the oocyte sperm previously incubated with foreign DNA. As semen samples collected in human infertility clinics are frequently contaminated with bacteria, one risk associated with the ICSI procedure is the injection of foreign, sperm-associated exogenous DNA into the oocyte, and the generation of transgenic offspring. Methods To analyse this possibility, ICSI was performed in mouse oocytes with frozen-thawed and Percoll-treated fresh sperm samples intentionally contaminated with plasmid EGFP-transformed E. coli bacteria or medium from which these bacteria were washed. Fertilized embryos were cultured in vitro until morula/blastocyst stage, transferred into pseudopregnant females, and at day 14, fetuses and reabsorptions were analysed by PCR for the genomic presence of integrated plasmid and/or bacterial DNA. Results Independently of the sperm pretreatment tested, transgenesis was produced. Conclusions Inadvertent transgenesis by conventional ICSI is a possibility that should not be neglected. Particular precautions, such as full bacteriological semen examinations and effective antibiotic semen pretreatments, should be taken in human infertility clinics, in order to exclude the possibility of accidental transgenesis. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
format journal article
author Moreira, P. N.
Fernández-González, R.
Rizos, D.
Ramirez, M.
Perez-Crespo, M.
Gutiérrez-Adán, A.
spellingShingle Moreira, P. N.
Fernández-González, R.
Rizos, D.
Ramirez, M.
Perez-Crespo, M.
Gutiérrez-Adán, A.
Inadvertent transgenesis by conventional ICSI in mice
author_facet Moreira, P. N.
Fernández-González, R.
Rizos, D.
Ramirez, M.
Perez-Crespo, M.
Gutiérrez-Adán, A.
author_sort Moreira, P. N.
title Inadvertent transgenesis by conventional ICSI in mice
title_short Inadvertent transgenesis by conventional ICSI in mice
title_full Inadvertent transgenesis by conventional ICSI in mice
title_fullStr Inadvertent transgenesis by conventional ICSI in mice
title_full_unstemmed Inadvertent transgenesis by conventional ICSI in mice
title_sort inadvertent transgenesis by conventional icsi in mice
publishDate 2005
url http://hdl.handle.net/20.500.12792/2990
work_keys_str_mv AT moreirapn inadvertenttransgenesisbyconventionalicsiinmice
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AT perezcrespom inadvertenttransgenesisbyconventionalicsiinmice
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