Animal Reproduction (AR)
Animal Reproduction (AR)

Alternative strategies for nuclear reprogramming in Somatic Cell Nuclear transfer (SCNT)

Pasqualino Loi, Domenico Iuso, Paola Toschi, Luca Palazzese, Marta Czernik

Downloads: 0
Views: 621


Twenty years passed by since the production of Dolly the sheep, but despite significant technical progress has been achieved in the manipulation procedures, the proportion of offspring following transfer of SCNT embryos has remained almost unchanged in farm animals. Remarkable progress has been obtained instead in laboratory animals, particularly by Japanese Groups, in the mouse. However, the nuclear reprogramming strategies tested in mouse do not always work in farm animals, and others are difficult to be implemented, for require complicated molecular biology tools unavailable yet in large animals. In this review we put in contest the previous work done in farm and laboratory animals with recent achievements obtained in our laboratory, and we also indicate a road map to increase the reliability of SCNT procedures.


somatic cell nuclear transfer, protamine 1, reprogramming, large animals.


Akama K, Kondo M, Sato H, Nakano M. 1999. Transition protein 4 from boar late spermatid nuclei is a topological factor that stimulates DNA-relaxing activity of topoisomerase I. FEBS Lett, 442:189-192.

Brock WA, Trostle PK, Meistrich ML. 1980. Meiotic synthesis of testis histones in the rat. Proc Natl Acad Sci. U S A, 77:371-375.

Chereji RV, Morozov AV. 2015. Functional roles of nucleosome stability and dynamics. Brief Funct Genomics, 14:50-60.

De Vries, M, Ramos L, Housein Z, De Boer P. 2012. Chromatin remodelling initiation during human spermiogenesis. Biol Open 1, 446-457.

Govin J, Khochbin S. 2013. Histone variants and sensing of chromatin functional states. Nucleus, 4:438-442.

Hosseini SM, Dufort I, Nieminen J, Moulavi F, Ghanaei HR, Hajian M, Jafarpour F, Forouzanfar M, Gourbai H, Shahverdi AH, Nasr-Esfahani MH, Sirard MA. 2016. Epigenetic modification with trichostatin A does not correct specific errors of somatic cell nuclear transfer at the transcriptomic level; highlighting the non-random nature of oocyte-mediated reprogramming errors. BMC Genomics, 4:17:16. doi: 10.1186/s12864-015-2264-z

Iuso D, Czernik M, Toschi P, Fidanza A, Zacchini F, Feil R, Curtet S, Buchou T, Shiota H, Khochbin S, Ptak GE, Loi P. 2015. Exogenous Expression of Human Protamine 1 (hPrm1) Remodels Fibroblast Nuclei into Spermatid-like Structures. Cell Rep, 13:1765-1771.

Johnson GD, Lalancette C, Linnemann AK, Leduc jF, Boissonneault G, Krawetz SA. 2011. The sperm nucleus: chromatin, RNA, and the nuclear matrix. Reproduction, 141:21-36.

Keefer CL. Artificial cloning of domestic animals. 2015. Proc Natl Acad Sci U S A, 112:8874-8878. Kishigami S, Mizutani E, Ohta H, Hikichi T, Thuan NV, Wakayama S, Bui HT, Wakayama T. 2006.

Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer. Biochem Biophys Res Commun, 340:183-189.

Kishigami S, Bui HT, Wakayama S, Tokunaga K, Van Thuan N, Hikichi T, Mizutani E, Ohta H, Suetsugu R, Sata T, Wakayama T. 2007. Successful mouse cloning of an outbred strain by trichostatin A treatment after somatic nuclear transfer. J Reprod Dev, 53:165-170.

Lewis JD, Saperas N, Song Y, Zamora MJ, Chiva M, Ausió J. 2004. Histone H1 and the origin of protamines. Proc Natl Acad Sci U S A, 101:4148-4152.

Loi P, Ptak G, Barboni B, Fulka J Jr, Cappai P, Clinton M. 2001. Genetic rescue of an endangered mammal by cross-species nuclear transfer using postmortem somatic cells. Nat Biotechnol, 19:962-964.

Loi P, Clinton M, Barboni B, Fulka J Jr, Cappai P, Feil R, Moor RM, Ptak G. 2002. Nuclei of nonviable ovine somatic cells develop into lambs after nuclear transplantation. Biol Reprod, 67:126-132.

Loi P, Clinton M, Vackova I, Fulka J Jr, Feil R, Palmieri C, Della Salda L, Ptak G. 2006. Placental abnormalities associated with post-natal mortality in sheep somatic cell clones. Theriogenology, 65:1110- 1121.

Loi P, Toschi P, Zacchini F, Ptak G, Scapolo PA, Capra E, Stella A, Marsan PA, Williams JL. 2016a. Synergies between assisted reproduction technologies and functional genomics. Genet Sel Evol, 48:53.

Loi P, Iuso D, Czernik M, Ogura A. 2016b. A New, Dynamic Era for Somatic Cell Nuclear Transfer? Trends Biotechnol, 34:791-797.

Loppin B, Bonnefoy E, Anselme C, Laurençon A, Karr TL, Couble P. 2005. The histone H3.3 chaperone HIRA is essential for chromatin assembly in the male pronucleus. Nature, 437:1386-1390.

Martínez-Soler F, Kurtz K, Ausió J, Chiva M. 2007. Transition of nuclear proteins and chromatin structure in spermiogenesis of Sepia officinalis. Mol Reprod Dev, 74:360-370.

Matoba S, Inoue K, Kohda T, Sugimoto M, Mizutani E, Ogonuki N, Nakamura T, Abe K, Nakano T, Ishino F, Ogura A. 2011. RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos. Proc Natl Acad Sci U S A, 108:20621-20626.

Matoba S, Liu Y, Lu F, Iwabuchi AK, Shen L, Inoue A, Zhang Y. 2014. Embryonic Development following Somatic Cell Nuclear Transfer Impeded by Persisting Histone Methylation. Cell, 159:884-895.

Miller D, Brinkworth M, Iles D. 2010. Paternal DNA packaging in spermatozoa: more than the sum of its parts? DNA, histones, protamines and epigenetics. Reproduction, 139:287-301.

Montellier E, Rousseaux S, Khochbin S. 2012. Crossfire over the nucleosome: molecular basis of postmeiotic male haploid genome compaction. Med Sci (Paris), 28:485-489.

Ogura, A., Inoue, K. and Wakayama, T. 2013. Recent advancements in cloning by somatic cell nuclear transfer. Philos Trans R Soc Lond B Biol Sci, 368:20110329.

Okuwaki M, Sumi A, Hisaoka M, SaotomeNakamura A, Akashi S, Nishimura Y, Nagata K. 2012. Function of homo- and hetero-oligomers of human nucleoplasmin/nucleophosmin family proteins NPM1, NPM2 and NPM3 during sperm chromatin remodelling. Nucleic Acids Res, 40:4861-4878.

Palmer DK, O'Day K, Margolis RL. 1990. The centromere specific histone CENP-A is selectively retained in discrete foci in mammalian sperm nuclei. Chromosoma, 100:32-36.

Pimm SL, Raven P. Biodiversity. 2000. Extinction by numbers. Nature, 24:843-845.

Pivot-Pajot C, Caron C, Govin J, Vion A, Rousseaux S, Khochbin S. 2003. Acetylation-dependent chromatin reorganization by BRDT, a testis-specific bromodomain-containing protein. Mol Cell Biol, 23:5354-5365.

Sangalli JR, De Bem TH, Perecin F, Chiaratti MR, Oliveira Lde J, de Araújo RR, Valim Pimentel JR, Smith LC, Meirelles FV. 2012. Treatment of nucleardonor cells or cloned zygotes with chromatin-modifying agents increases histone acetylation but does not improve full-term development of cloned cattle. Cell Reprogram, 14:235-247

Saragusty J, Diecke S, Drukker M, Durrant B, Friedrich Ben-Nun I, Galli C, Göritz F, Hayashi K, Hermes R, Holtze S, Johnson S, Lazzari G, Loi P, Loring JF, Okita K, Renfree MB, Seet S, Voracek T, Stejskal J, Ryder OA. 2016. Hildebrandt TB Rewinding the process of mammalian extinction. Zoo Biol, 35:280-292.

Singh J, Rao MR. 1988. Interaction of rat testis protein TP, with nucleosome core particle. Biochem Int, 17:701-710.

Taylor-Robinson AW, Walton S, Swain DL, Walsh KB, Vajta G. 2014. The potential for modification in cloning and vitrification technology to enhance genetic progress in beef cattle in Northern Australia. Anim Reprod Sci, 148:91-96.

Van Thuan N, Bui HT, Kim JH, Hikichi T, Wakayama S, Kishigami S, Mizutani E, Wakayama T. 2009. The histone deacetylase inhibitor scriptaid enhances nascent mRNA production and rescues fullterm development in cloned inbred mice. Reproduction, 138:309-317.

Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH. 1997. Viable offspring derived from fetal and adult mammalian cells. Nature, 385:810-813.

Wu F, Caron C, De Robertis C, Khochbin S, Rousseaux, S. 2008. Testis-specific histone variants H2AL1/2 rapidly disappear from paternal heterochromatin after fertilization. J Reprod Dev, 54:413-417.

5b7192ff0e88253f4a8068a8 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections