The Generation of Cloned Drosophila melanogaster

Haigh, Andrew; MacDonald, William; Lloyd, Vett K.

doi:10.1534/genetics.104.035113

Cited by 10 publications

(9 citation statements)

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“…However, cloning procedures have been recently adapted to very unconventional animal models with two papers reporting about successful cloning of zebra fish (Ju et al. 2004) and Drosophila (Haigh et al. 2005) these accomplishments broad the range of species that can potentially benefit from cloning technology.…”

Section: Discussionmentioning

confidence: 99%

Nuclear Transfer of Freeze‐Dried Somatic Cells into Enucleated Sheep Oocytes

Loi

Matzukawa

Ptak

et al. 2008

Reprod Domestic Animals

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Lyophilization has been used since long time to preserve yeast and bacteria strains. Subsequently, a great deal of efforts has been dedicated to the preservation in a dry state of red blood cells and platelets. However, despite more than 30 years passed by, no significant progress has been achieved. Recently, it has been reported that freeze-dried mice spermatozoa were able to generate normal offspring following injection into the mature mice oocytes. In this work, we prompted to apply the lyophilization protocol developed for mice spermatozoa to sheep somatic cells (lymphocytes and granulosa cells). More than 350 enucleated sheep oocytes were injected with granulosa cells, and freeze dried using the protocol developed for mice sperm cells. Transplanted nuclei organized large pronuclei with fragmented DNA, but none of them entered the first mitosis. In the second part of the experiments, trehalose and EGTA were found to reduce significantly the extent of nuclear damage (65% and 55% intact nuclei in lymphocyte and granulosa cells, respectively) following freeze drying. Granulosa cells lyophilized with EGTA/trehalose and stored at room temperature for 3 years were used for nuclear transfer, and the injected oocytes were cultured in vitro for 7 days. Approximately 16% of the oocyte injected with freeze-dried cells developed into blastocysts. To conclude, we demonstrated for the first time that nucleated cells maintain genomic integrity after prolonged storage in a dry state, and we were able to achieve early embryonic development following injection of these cells into enucleated sheep oocytes.

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Section: Discussionmentioning

confidence: 99%

Nuclear Transfer of Freeze‐Dried Somatic Cells into Enucleated Sheep Oocytes

Loi

Matzukawa

Ptak

et al. 2008

Reprod Domestic Animals

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“…Likewise, about 97% Drosophila transplant recipients die during embryogenesis and of those that do hatch, only approximately one-third eclose as adults. 23 Gross morphological defects such as tracheal disorganization and the absence of anterior structures are observed in many cloned embryos and larvae. 23 The underlying cause of these defects is not yet known.…”

Section: The Trouble With Clones-epigenetic Defects In Cloned Animalsmentioning

confidence: 99%

“…23 Gross morphological defects such as tracheal disorganization and the absence of anterior structures are observed in many cloned embryos and larvae. 23 The underlying cause of these defects is not yet known. Both mammalian and insect clone survival is adversely affected by a number of factors, including mechanical damage to the embryo, discordant donor and recipient cell cycle and disrupted epigenetic gene regulation.…”

Section: The Trouble With Clones-epigenetic Defects In Cloned Animalsmentioning

confidence: 99%

“…In 2005, we added Drosophila to the list of organisms generated by nuclear transplantation. 23 We subsequently showed that the epigenetic silencing of an imprinted gene is lost in cloned Drosophila, 24 as is true of many cloned mammals. The ability to produce cloned Drosophila, coupled with the sophisticated understanding of early development and the large repertoire of mutations in genes regulating epigenetic silencing 25,2,27 make Drosophila an ideal model system in which to study, and potentially mitigate, the epigenetic defects resulting from nuclear transplantation.…”

Section: Introductionmentioning

confidence: 99%

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Why clone flies? Using ClonedDrosophilato Monitor Epigenetic Defects

Haigh

Lloyd²

2007

Fly

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Since the birth of the first cloned sheep in 1996, advances in nuclear transplantation have led to both the creation of genetically tailored stem cells and the generation of a number of cloned organisms. The list of cloned animals reared to adulthood currently includes the frog, sheep, mouse, cow, goat, pig, rabbit, cat, zebrafish, mule, horse, rat and dog. The addition of Drosophila to this elite bestiary of cloned animals has prompted the question - why clone flies? Organisms generated by nuclear transplantation suffer from a high rate of associated defects, and many of these defects appear to be related to aberrant genomic imprinting. Imprinted gene expression also appears to be compromised in Drosophila clones. Proper imprinted gene regulation relies on a suite of highly conserved chromatin-modifying genes first identified in Drosophila. Thus, Drosophila can potentially be used to study epigenetic dysfunction in cloned animals and to screen for genetic and epigenetic conditions that promote the production of healthy clones.

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“…We believe Drosophila would be excellent host for our massively parallel automation experiment. Haigh et al [25] have successfully used the embryonic nuclear transfer to create viable adult Drosophila clones and approximately 10% of recipients survive through embryogenesis, while less than 1% eclose and metamorphose as viable adults. We wish to increase the rate of successful cloning with the tools of bio-NanoRobot and 3-D microfluidics system.…”

Section: Figurementioning

confidence: 99%

A bio-NanoRobot design for drosophila therapeutic cloning

Chang

Szu

2006

Independent Component Analyses, Wavelets, Unsupervised Smart Sensors, and Neural Networks IV

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To investigate Somatic Cell Nuclear Transfer (SCNT), we choose the Drosophila cloning based on a recent experiment (Haigh, MacDonald, Lioyd, Gen. V.169,1165 to be improving the adulthood rate in 2-week turn-around time.Original 1% success rate might be due to three less certain key steps: (i) The double membranes of a nucleus has at its pore led to the attached Rough Endoplasmic Reticulum (ER), passing the genetic instruction to assemble amino acids, proteins and lipid at its smooth end. Also, any mismatch of nucleus with mitochondria (MT) having own small genome for energy production had led to reprogramming failure. (D. Wallace, UC Irvine, Nature,Vol. 439, pp.653). We ask "whether a guest DNA shall come with its servants, ER, MT, etc or not." It seemed to be logical to have a whole package replaced the embryonic host cell, equipped with all housekeeping, energy production and mitosis functionalities except the genetic information. To answer this hypothesis, we design a bio-NanoRobot having a surgical precision in removing the desired nucleus with or without its attached ER and MT material. The design is based on a real-time multiplexing principle of combining both the soft-contact-vision of the Nobel Laureate Binning called Atomic Force Microscope (AFM) and the hard-grasp-action called NanoRobot TM by Xi and Szu, 2004. However, applying it, we must re-design a new bio-NanoRobot, consisting of two parts: (a) multiple resolution analysis (MRA) using AI to control a dual-resolution vision system: the soft-contact-vision AFM co-registered with a on-contact high resolution imaging; and (b) two cantilever arms capable to hold and enucleate a cell. The calibration and automation are controlled by AI Case-Based reasoning (CBR) together with AI Blackboard (BB) of the taxonomy, necessary for integrating different tool's tolerance and resolution at the same location. Moreover, keeping the biological sample in one place, while a set of tools rotates upon it similar to a set of microscopic lenses, we can avoid the non-real-time re-imaging, and inadvertent contamination. Applying an imposing electrical field, we can take the advantage of structure differences in smooth nuclear membranes inducing Van der Waal's forces versus random cytoplasm. (ii) The re-programming of transplanted cells to the ground state is unclear and usually relies on electrochemical means tested systematically in a modified 3D Caltech micro-fluidics. (iii) Our real-time MRA video-manipulator can elucidate the mitosis's tread-mill assembly mechanism in the development course of pluripotent stem cell differentiation into specialized tissue cell engineering. Such a combination bio-NanoRobot and micro-fluidic massive parallel assembly-line approach might not only replace the aspirating pipette with a self-enucleating Drosophila embryonic eggs, but also genetically reproduce a large amount of cloning embryonic eggs repeatedly for various re-programming hypotheses.

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The Generation of Cloned Drosophila melanogaster

Cited by 10 publications

References 9 publications

Nuclear Transfer of Freeze‐Dried Somatic Cells into Enucleated Sheep Oocytes

Nuclear Transfer of Freeze‐Dried Somatic Cells into Enucleated Sheep Oocytes

Why clone flies? Using ClonedDrosophilato Monitor Epigenetic Defects

A bio-NanoRobot design for drosophila therapeutic cloning

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