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Generation of Yeast Artificial Chromosome Transgenic Mice by Intracytoplasmic Sperm Injection

AutorMoreira, Pedro N.; Pozueta, Julio; Giraldo, Patricia; Gutiérrez-Adán, Alfonso; Montoliu, Lluís
Palabras claveArtificial chromosomes
Intracytoplasmic sperm injection (ICSI)
In vitro fertilization
Sperm-mediated gene transfer (SMGT)
Transgene integrity
Artificial chromosomes
Assisted reproductive technology (ART)
Fecha de publicación26-oct-2007
EditorHumana Press
CitaciónIn: Methods in Molecular Biology, vol. 349; YAC Protocols: Second Edition, 151-161
ResumenGenomic-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 pronuclear 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.
Descripción11 pages, 1 figure.-- PMID: 17071981 [PubMed].
Versión del editorhttp://dx.doi.org/10.1385/1-59745-158-4:151
ISBN978-1-58829-612-2 (Print)
978-1-59745-158-1 (Online)
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