Construction of a swine artificial chromosome: a novel vector for transgenesis in the pig

Poggiali, Paola; Scoarughi, Gian Luca; Lavitrano, Marialuisa; Donini, Pierluigi; Cimmino, Carmen

doi:10.1016/s0300-9084(02)00019-6

Cited by 8 publications

(12 citation statements)

References 19 publications

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“…The DNA sequences considered in this study were the swine centromeric sequences previously used in SAC construction (accession nos. X62138.1, X62139.1 and X62140.1; Poggiali et al 2002); a human alphasatellite sequence (accession no. AJ717298); sheep satellite I and satellite II sequences (accession nos.…”

Section: Methodsmentioning

confidence: 99%

“…YAC d1Neo2 gave rise to the SAC d1Neo2 in PK15 swine cells (Poggiali et al 2002). The YAC is maintained in d1 by growth on Ymin medium lacking uracil and lysine.…”

Section: Methodsmentioning

confidence: 99%

“…Cultured cells grown to 60% confluence in 100 mm plates were incubated in fresh medium containing 2 μg/ml colcemid (Roche, Mannheim, Germany) for 4 h and then treated according to standard procedures with 0.075 M KCl and fixed in methanol-acetic acid (3:1). Artificial chromosomes were detected in the presence of 15 μg/μl of salmon sperm DNA with a biotin-labelled pCG probe obtained by nick translation (pCGS990meganeo probe of Poggiali et al 2002) in both FLK clones and PK15 control clones. This probe is specific for the MAC and corresponds to the left arm of the original YAC d1Neo2.…”

Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%

“…The DNA sequences considered were the swine centromeric sequences previously used for the construction of a SAC (Poggiali et al 2002), a human alpha-satellite sequence, sheep satellite I and II sequences, a bovine satellite DNA sequence, and the mouse minor satellite sequence. These sequences were compared using the local alignment algorithm LALIGN: in the case of repetitive sequences it is appropriate to use a strategy that compares them pairwise and locates local alignment.…”

Section: Mammalian Centromeric Sequence Homologymentioning

confidence: 99%

“…On the basis of the centromeric sequence homology results obtained we decided to transfer into fetal lamb kidney cells (FLK) the YAC precursor of the swine MAC d1Neo2 previously assembled by us and used to generate a SAC (Poggiali et al 2002). In this manner the ability of swine centromeric sequences to drive MAC formation in a non-swine mammalian cell line could be evaluated.…”

Section: Sac Precursor Transfer Into Sheep Cellsmentioning

confidence: 99%

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Interspecific transfer of mammalian artificial chromosomes between farm animals

2009

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It is often desirable to transfer a mammalian artificial chromosome (MAC) from the cells of one species to those of another. Attempts to carry out such transfer have been successful in some cases and have failed in others. In this study we have tested the hypothesis that centromeric DNA sequence similarity could be a useful criterion for determining MAC host range. Homology studies indicated that the sheep should give positive transfer results. The prediction was tested by introducing into sheep cells a yeast artificial chromosome that contained swine centromeric sequences and that had previously been used to produce a de novo MAC in swine cells. The experiments resulted in the formation of a functional de novo MAC in sheep cells, as attested by FISH analysis. The newly formed MAC remained structurally and functionally stable in ovine up to 52 generations. The centromeric sequences present on the newly formed MAC are probably swine sequences, although it cannot be ruled out that some sheep sequences may also have migrated to the MAC. The size of the sheep MAC was determined by atomic force microscopy. Thus, centromeric sequence similarity appears to be a useful criterion for predicting the animal species between which MACs can shuttle.

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Section: Methodsmentioning

confidence: 99%

“…YAC d1Neo2 gave rise to the SAC d1Neo2 in PK15 swine cells (Poggiali et al 2002). The YAC is maintained in d1 by growth on Ymin medium lacking uracil and lysine.…”

Section: Methodsmentioning

confidence: 99%

Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%

Section: Mammalian Centromeric Sequence Homologymentioning

confidence: 99%

Section: Sac Precursor Transfer Into Sheep Cellsmentioning

confidence: 99%

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Interspecific transfer of mammalian artificial chromosomes between farm animals

2009

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Yeast Artificial Chromosomes

Bruschi

Gjuračić

Tosato

2012

Encyclopedia of Life Sciences

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Yeast artificial chromosomes (YACs) are shuttle‐vectors that can be amplified in bacteria and employed for the cloning and manipulation of large deoxyribonucleic acid (DNA) inserts (up to 3 Mb pairs) in the yeast Saccharomyces cerevisiae . Artificial chromosomes can be conveniently built and modified in yeast cells using in vivo homologous recombination, a novel process known as ‘recombineering’. The capacity of YACs to accommodate large DNA fragments is exploited to clone clusters of genes surrounded by their native DNA context, where regulatory elements are located. This is important for biotechnology, when YACs are used for engineering genetic determinants of new biochemical pathways for production of secondary metabolites and for heterologous protein expression. YACs can be retrofitted with the appropriate selectable markers and transmitted to cells of different organisms allowing the generation of transgenic animals. Finally, YACs are largely employed in the production of full‐scale genomic libraries, for mapping and functional analysis purposes. Key Concepts: Yeast artificial chromosomes (YACs) represent the top instruments for the study of eukaryotic genomes and for mobilisation of large genetic elements among bacteria and eukaryotes. YACs behave like naturally existing chromosomes, provided that they are of the proper size, showing comparable stability. YACs can be manipulated directly by classical genetic engineering as well as by modern recombineering technology. Properly retrofitted, YACs can be used in many different organisms, for cloning or genome analysis. Chromosomal translocation can be studied using disposable YACs that do not harbour genetic information essential for cell function.

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Nuclear Remodeling and Nuclear Reprogramming for Making Transgenic Pigs by Nuclear Transfer

Prather

Somatic Cell Nuclear Transfer

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A better understanding of the cellular and molecular events that occur when a nucleus is transferred to the cytoplasm of an oocyte will permit the development of improved procedures for performing nuclear transfer and cloning. In some cases it appears that the gene(s) are reprogrammed, while in other cases there appears to be little effect on gene expression. Not only does the pattern of gene expression need to be reprogrammed, but other structures within the nucleus also need to be remodeled. While nuclear transfer works and transgenic and knockout animals can be created, it still is an inefficient process. However, even with the current low efficiencies this technique has proved very valuable for the production of animals that might be useful for tissue or organ transplantation to humans.

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Construction of a swine artificial chromosome: a novel vector for transgenesis in the pig

Cited by 8 publications

References 19 publications

Interspecific transfer of mammalian artificial chromosomes between farm animals

Interspecific transfer of mammalian artificial chromosomes between farm animals

Yeast Artificial Chromosomes

Nuclear Remodeling and Nuclear Reprogramming for Making Transgenic Pigs by Nuclear Transfer

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