Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection
Genomic-type transgenes are usually expressed in appropriate spatial- and temporal-specific manners. The largest genomic transgenes can be prepared using yeast artificial chromosomes (YACs). Normally, YAC transgenic mice are produced by standard pro-nuclear microinjection, although other methods, involving the use of embryonic stem (ES) cells, have been also devised. To overcome the difficulty and time extension of ES cell-type approaches and to improve the rather usual low efficiency of YAC DNA transgenesis by pronuclear microinjection, that is mostly dependent on the YAC DNA quality of samples, we have devised an updated intracytoplasmic sperm injection (ICSI) method for the stable incorporation of YACs into the germ line of mice. DNA transgenesis efficiencies achieved are often 10 times greater than those usually obtained by standard microinjection, thus enabling the identification of either more transgenic founder animals and the use of reduced numbers of individuals in animal experimentation.
Main Authors: | , , , , |
---|---|
Format: | artículo biblioteca |
Language: | English |
Published: |
Springer
2006
|
Subjects: | Artificial chromosomes, Intracytoplasmic sperm injection (ICSI), In vitro fertilization, Sperm-mediated gene transfer (SMGT), Transgene integrity, Assisted reproductive technology (ART), |
Online Access: | http://hdl.handle.net/20.500.12792/2994 http://hdl.handle.net/10261/292551 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
dig-inia-es-10261-292551 |
---|---|
record_format |
koha |
spelling |
dig-inia-es-10261-2925512023-02-20T07:30:05Z Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection Moreira, P. N. Pozueta, J. Giraldo, P. Gutiérrez Adán, Alfonso Montoliu, L. Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) Genomic-type transgenes are usually expressed in appropriate spatial- and temporal-specific manners. The largest genomic transgenes can be prepared using yeast artificial chromosomes (YACs). Normally, YAC transgenic mice are produced by standard pro-nuclear microinjection, although other methods, involving the use of embryonic stem (ES) cells, have been also devised. To overcome the difficulty and time extension of ES cell-type approaches and to improve the rather usual low efficiency of YAC DNA transgenesis by pronuclear microinjection, that is mostly dependent on the YAC DNA quality of samples, we have devised an updated intracytoplasmic sperm injection (ICSI) method for the stable incorporation of YACs into the germ line of mice. DNA transgenesis efficiencies achieved are often 10 times greater than those usually obtained by standard microinjection, thus enabling the identification of either more transgenic founder animals and the use of reduced numbers of individuals in animal experimentation. 2023-02-20T07:30:05Z 2023-02-20T07:30:05Z 2006 artículo Methods in Molecular Biology 349: 151-161 (2006) 1064-3745 http://hdl.handle.net/20.500.12792/2994 http://hdl.handle.net/10261/292551 1940-6029 en none Springer |
institution |
INIA ES |
collection |
DSpace |
country |
España |
countrycode |
ES |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-inia-es |
tag |
biblioteca |
region |
Europa del Sur |
libraryname |
Biblioteca del INIA España |
language |
English |
topic |
Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) |
spellingShingle |
Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) Moreira, P. N. Pozueta, J. Giraldo, P. Gutiérrez Adán, Alfonso Montoliu, L. Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
description |
Genomic-type transgenes are usually expressed in appropriate spatial- and temporal-specific manners. The largest genomic transgenes can be prepared using yeast artificial chromosomes (YACs). Normally, YAC transgenic mice are produced by standard pro-nuclear microinjection, although other methods, involving the use of embryonic stem (ES) cells, have been also devised. To overcome the difficulty and time extension of ES cell-type approaches and to improve the rather usual low efficiency of YAC DNA transgenesis by pronuclear microinjection, that is mostly dependent on the YAC DNA quality of samples, we have devised an updated intracytoplasmic sperm injection (ICSI) method for the stable incorporation of YACs into the germ line of mice. DNA transgenesis efficiencies achieved are often 10 times greater than those usually obtained by standard microinjection, thus enabling the identification of either more transgenic founder animals and the use of reduced numbers of individuals in animal experimentation. |
format |
artículo |
topic_facet |
Artificial chromosomes Intracytoplasmic sperm injection (ICSI) In vitro fertilization Sperm-mediated gene transfer (SMGT) Transgene integrity Assisted reproductive technology (ART) |
author |
Moreira, P. N. Pozueta, J. Giraldo, P. Gutiérrez Adán, Alfonso Montoliu, L. |
author_facet |
Moreira, P. N. Pozueta, J. Giraldo, P. Gutiérrez Adán, Alfonso Montoliu, L. |
author_sort |
Moreira, P. N. |
title |
Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
title_short |
Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
title_full |
Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
title_fullStr |
Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
title_full_unstemmed |
Generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
title_sort |
generation of yeast artificial chromosome transgenic mice by intracytoplasmic sperm injection |
publisher |
Springer |
publishDate |
2006 |
url |
http://hdl.handle.net/20.500.12792/2994 http://hdl.handle.net/10261/292551 |
work_keys_str_mv |
AT moreirapn generationofyeastartificialchromosometransgenicmicebyintracytoplasmicsperminjection AT pozuetaj generationofyeastartificialchromosometransgenicmicebyintracytoplasmicsperminjection AT giraldop generationofyeastartificialchromosometransgenicmicebyintracytoplasmicsperminjection AT gutierrezadanalfonso generationofyeastartificialchromosometransgenicmicebyintracytoplasmicsperminjection AT montoliul generationofyeastartificialchromosometransgenicmicebyintracytoplasmicsperminjection |
_version_ |
1767603360623493120 |