The Mechanism of Binding of Exogenous DNA to Sperm Cells: Factors Controlling the DNA Uptake

Zani, Massimo; Lavitrano, Marialuisa; French, Deborah; Lulli, Valentina; Maione, Barbara; Sperandio, Sabina; Spadafora, Corrado

doi:10.1006/excr.1995.1063

Cited by 119 publications

(88 citation statements)

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“…The exogenous DNA interacts with DNAbinding proteins (DBPs) of 30-35 kDa, which are present on the sperm cell surface (Lavitrano et al 1992;Zani et al 1995) and we have demonstrated that major histocompatibility complex (MHC) class II and CD4 molecules also play a role in the process of sperm/DNA interaction (Lavitrano et al 1997b). In fact, sperm cells from MHC class II-knockout mice have a reduced ability to bind DNA compared with sperm cells from wild-type animals Wu et al 1990).…”

Section: Molecular Mechanism Of Sperm-mediated Gene Transfermentioning

confidence: 99%

“…This factor also exerted a powerful inhibitory effect on DNA uptake in sperm cells of heterologous species. The DBPs appeared to be the specific target through which the inhibition is mediated because, in the presence of the inhibitory factor, the DBPs lose their ability to bind exogenous DNA (Zani et al 1995). Thus, the interaction of exogenous DNA with sperm cells does not appear to be a casual event but, in contrast, relies on a molecular mechanism based on the cooperation of specific protein factors.…”

Section: Molecular Mechanism Of Sperm-mediated Gene Transfermentioning

confidence: 99%

“…In 1989, we described a new method for the production of transgenic animals, sperm-mediated gene transfer (SMGT), which it is based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA and to transfer it into the egg at fertilisation (Lavitrano et al 1989(Lavitrano et al , 1992(Lavitrano et al , 1997bFrancolini et al 1993;Zani et al 1995), as illustrated in Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

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Sperm-mediated gene transfer

Lavitrano

Busnelli

Cerrito

et al. 2006

Reprod. Fertil. Dev.

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115

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Abstract. Since 1989, a new method for the production of transgenic animals has been available, namely spermmediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in the mouse. In addition, we successfully adapted and optimised the technique for use in large animals; it was, in fact, highly efficient in the generation of human decay accelerating factor transgenic pig lines, as well as multigene transgenic pigs in which three different reporter genes, namely enhanced green fluorescent protein, enhanced blue fluorescent protein and red fluorescent protein, were introduced. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared with other methods. Furthermore, SMGT does not require embryo handling or expensive equipment. Sperm-mediated gene transfer could also be used to generate multigene transgenic pigs that would be of benefit as large animal models for medical research, for agricultural and pharmaceutical applications and, in particular, for xenotransplantation, which requires extensive genetic manipulation of donor pigs to make them suitable for grafting to humans.

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Section: Molecular Mechanism Of Sperm-mediated Gene Transfermentioning

confidence: 99%

Section: Molecular Mechanism Of Sperm-mediated Gene Transfermentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Sperm-mediated gene transfer

Lavitrano

Busnelli

Cerrito

et al. 2006

Reprod. Fertil. Dev.

Self Cite

115

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“…In this process, binding of exogenous DNA is thought to involve sperm submembrane compartment proteins and occurs in demembranated, Triton X-100-treated sperm heads (66). Specific localization of exogenous DNA binding to sperm appears confined to PT structures, such as the subacrosomal layer and the post-acrosomal sheath, and may involve proteins of less than 20 kDa in sperm of various species (67). Given the localization of PT-derived somatic histones characterized in this study, and the inherent capabilities of somatic histones for DNA binding, it is likely that they are potential candidates for this process.…”

Section: Figmentioning

confidence: 99%

Somatic Histones Are Components of the Perinuclear Theca in Bovine Spermatozoa

Tovich

Oko

2003

Journal of Biological Chemistry

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The perinuclear theca is a non-ionic detergent-resistant, electron-dense layer surrounding the condensed nucleus of mammalian sperm. The known proteins originating from the perinuclear theca have implicated the structure in a variety of important cellular processes during spermiogenesis and fertilization. Nonetheless, the composition of the perinuclear theca remains largely unexplored. We have isolated a group of low molecular mass (14 -19 kDa) perinuclear theca-derived proteins from acrosome-depleted bovine sperm heads by salt (1 M KCl) extraction and have identified them as core somatic histones. N-terminal sequencing and immunoblotting with anti-histone antibodies confirmed the presence of both intact and proteolytically cleaved somatic histones H3, H2B, H2A, and H4. Identical proteins were isolated using 2% SDS or 1 N HCl extractions. Subsequent acid and SDS extractions of intact bovine sperm revealed the presence of all four intact histone subtypes, with minimal proteolysis. Two-dimensional acid/urea/Triton-SDS-PAGE, coupled with immunoblotting analysis, confirmed the somatic nature of these perinuclear theca-derived histones. Estimates of the abundance of perinuclear theca-derived histones showed that up to 0.2 pg per sperm of each histone subtype was present. Immunogold labeling at the ultrastructural level localized all four core somatic histones to the postacrosomal sheath region of bovine epididymal sperm, when probed with affinity-purified anti-histone antibodies. Little immunoreactivity was detected in residual perinuclear theca structures following the extractions. Taken together, these findings indicate the unprecedented and stable localization of non-nuclear somatic histones in bovine sperm perinuclear theca.The perinuclear theca (PT) 1 is a specialized cytoskeletal component residing under the acrosome and almost completely surrounding the nucleus of mammalian sperm heads. Based on morphological and protein composition assessments, the PT is divided into two structurally continuous regions, termed the subacrosomal layer, and the post-acrosomal sheath (reviewed in Refs. 1 and 2). Although numerous proteins have been characterized from these PT regions (reviewed in Ref. 3), few of the known PT proteins to date resemble traditional cytoskeletal proteins. Rather, the PT contains a range of proteins of varying function, including Calicin, a basic structural protein that binds actin (4, 5); Stat4, a transcriptional activator hypothesized to contribute to zygotic gene activation (6); SubH2Bv (PT15), a histone H2B-like variant proposed to contribute to acrosome-nuclear docking (3); and PERF 15, a fatty acid-binding protein predominating in the perforatorium of murid sperm (7-9). The PT structure has been implicated in a number of essential cellular processes, such as nuclear shaping during spermiogenesis (2, 10), and egg activation and pronuclear formation during fertilization (11-15), although the PT proteins involved in these processes remain elusive. Their identification and characterization is theref...

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“…Sperm mediated gene transfer (SMGT) is an alternative technique using natural ability of spermatozoa as a vector to transfer exogenous DNA into the egg at fertilization (Bacci, 2007;Lavitranoet al, 2002 andZani, et al, 1995). This technique was first introduced in mice by Dr. Lavitrano in 1989.…”

Section: Sperm Mediated Gene Transfer (Smgt)mentioning

confidence: 99%

Review on current application of genomic biotechnologies to improve livestock and livestock products

2017

Int. J. Adv. Res. Biol. Sci

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An organism's genome is the 'blueprint of life,' the complete set of instructions for making the molecules, cells and tissues that control how an animal can perform throughout its life. Every cell in the body contains the complete genome, encoded in the form of DNA. This review was conducted to give emphasis about the role or the current application of some advance genomic biotechnologies in animal breeding, disease diagnosis and prevention and control strategies. There are opportunities for using molecular genetics to identify genes that are involved in variety of traits. Revolutionary opportunities for the modification of animal performance are being created by the development of new methods (reproductive technology) for embryo manipulation and the application of molecular biology (cloning and transgenesis). Consequently, new approaches are needed to develop improved tools and strategies for prevention and control of infectious diseases in animal agriculture. Among the most effective and successful of these tools are animal vaccines using recombinant deoxyribonucleic acid (rDNA) technologies. In the recent years a profound change has occurred with the introduction of new genomic biotechnological assays for the diagnosis of infectious diseases of livestock and zoonotic pathogens. These new assays include various forms of PCR, genomic sequencing, DNA probes and DNA microarray technology, Nanotechnology, Restriction fragment length polymorphisms and Pulsed field gel electrophoresis. Generally animal genomics is of interest because of its importance to produce high quality food products economically and efficiently to furnish for the increasing supply demand gap all over the world.

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The Mechanism of Binding of Exogenous DNA to Sperm Cells: Factors Controlling the DNA Uptake

Cited by 119 publications

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Sperm-mediated gene transfer

Sperm-mediated gene transfer

Somatic Histones Are Components of the Perinuclear Theca in Bovine Spermatozoa

Review on current application of genomic biotechnologies to improve livestock and livestock products

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