Showing papers by "Michele Morgante published in 2017"

Regeneration of the entire human epidermis using transgenic stem cells

Abstract: Junctional epidermolysis bullosa (JEB) is a severe and often lethal genetic disease caused by mutations in genes encoding the basement membrane component laminin-332. Surviving patients with JEB develop chronic wounds to the skin and mucosa, which impair their quality of life and lead to skin cancer. Here we show that autologous transgenic keratinocyte cultures regenerated an entire, fully functional epidermis on a seven-year-old child suffering from a devastating, life-threatening form of JEB. The proviral integration pattern was maintained in vivo and epidermal renewal did not cause any clonal selection. Clonal tracing showed that the human epidermis is sustained not by equipotent progenitors, but by a limited number of long-lived stem cells, detected as holoclones, that can extensively self-renew in vitro and in vivo and produce progenitors that replenish terminally differentiated keratinocytes. This study provides a blueprint that can be applied to other stem cell-mediated combined ex vivo cell and gene therapies. Autologous transgenic epidermal stem cell cultures are used to reconstitute almost the entire epidermis of a patient with severe junctional epidermolysis bullosa. Patients with junctional epidermolysis bullosa (JEB) carry mutations in genes that encode components of the basement membrane, which ensures the integrity between the epidermis and the dermis, such as laminin-332. These mutations cause blistering of the skin and chronic wounds. Following initial treatment of an adult patient with a limited affected region, Michele De Luca and colleagues reconstruct the full epidermis of a 7-year-old patient with autologous transgenic cells transduced with a virus vector carrying the non-mutated form of laminin-322. The integration sites of the virus used for gene delivery provide a tracing tool ex vivo and in vivo and demonstrate that the human epidermis is sustained by a limited number of long-lived stem cells.

A new version of the grapevine reference genome assembly (12X.v2) and of its annotation (VCost.v3).

Abstract: :The dataset comprises 12X.v2 grapevine reference genome assembly and its VCost.v3 gene annotation in Genbank format. The 12X.v2 chromosome assembly is based on the scaffolds of the grapevine reference genome build (FN594950-FN597014, EMBL release 102; Vitis vinifera cv. PN40024). The data set is provided in support of the data paper: “A new version of the grapevine reference genome assembly (12X.v2) and of its annotation (VCost.v3)” by Canaguier A, Grimplet J, Di Gaspero G, Scalabrin S, Duchene E, Mohellibi N, Guichard C, Choisne N, Rombault S, Le Clainche I, Berard A, Chauveau A, Bounon R, Ruztenholtz C, Morgante M, Le Paslier M-C, Brunel D, Adam-Blondon A-F (Genomic Data, 2017, https://doi.org/10.1016/j.gdata.2017.09.002)

A PLAG1 mutation contributed to stature recovery in modern cattle

Abstract: The recent evolution of cattle is marked by fluctuations in body size. Height in the Bos taurus lineage was reduced by a factor of ~1.5 from the Neolithic to the Middle Ages, and increased again only during the Early Modern Ages. Using haplotype analysis, we found evidence that the bovine PLAG1 mutation (Q) with major effects on body size, weight and reproduction is a >1,000 years old derived allele that increased rapidly in frequency in Northwestern European B. taurus between the 16th and 18th centuries. Towards the 19th and 20th centuries, Q was introgressed into non-European B. taurus and Bos indicus breeds. These data implicate a major role of Q in recent changes in body size in modern cattle, and represent one of the first examples of a genomic sweep in livestock that was driven by selection on a complex trait.

De novo assembly, functional annotation, and analysis of the giant reed ( Arundo donax L.) leaf transcriptome provide tools for the development of a biofuel feedstock

Abstract: Arundo donax has attracted renewed interest as a potential candidate energy crop for use in biomass-to-liquid fuel conversion processes and biorefineries. This is due to its high productivity, adaptability to marginal land conditions, and suitability for biofuel and biomaterial production. Despite its importance, the genomic resources currently available for supporting the improvement of this species are still limited. We used RNA sequencing (RNA-Seq) to de novo assemble and characterize the A. donax leaf transcriptome. The sequencing generated 1249 million clean reads that were assembled using single-k-mer and multi-k-mer approaches into 62,596 unique sequences (unitranscripts) with an N50 of 1134 bp. TransDecoder and Trinotate software suites were used to obtain putative coding sequences and annotate them by mapping to UniProtKB/Swiss-Prot and UniRef90 databases, searching for known transcripts, proteins, protein domains, and signal peptides. Furthermore, the unitranscripts were annotated by mapping them to the NCBI non-redundant, GO and KEGG pathway databases using Blast2GO. The transcriptome was also characterized by BLAST searches to investigate homologous transcripts of key genes involved in important metabolic pathways, such as lignin, cellulose, purine, and thiamine biosynthesis and carbon fixation. Moreover, a set of homologous transcripts of key genes involved in stomatal development and of genes coding for stress-associated proteins (SAPs) were identified. Additionally, 8364 simple sequence repeat (SSR) markers were identified and surveyed. SSRs appeared more abundant in non-coding regions (63.18%) than in coding regions (36.82%). This SSR dataset represents the first marker catalogue of A. donax. 53 SSRs (PolySSRs) were then predicted to be polymorphic between ecotype-specific assemblies, suggesting genetic variability in the studied ecotypes. This study provides the first publicly available leaf transcriptome for the A. donax bioenergy crop. The functional annotation and characterization of the transcriptome will be highly useful for providing insight into the molecular mechanisms underlying its extreme adaptability. The identification of homologous transcripts involved in key metabolic pathways offers a platform for directing future efforts in genetic improvement of this species. Finally, the identified SSRs will facilitate the harnessing of untapped genetic diversity. This transcriptome should be of value to ongoing functional genomics and genetic studies in this crop of paramount economic importance.

Reduction of heterozygosity (ROH) as a method to detect mosaic structural variation.

Abstract: Dipartimento di Scienze agroalimentari, ambientali e animali, Universit a di Udine, Udine, Italy Istituto di Genomica Applicata (IGA), Udine, Italy IGA Technology Services, Udine, Italy Parco Tecnologico Padano, Lodi, Italy Institute of Biosciences and Bioresources, National Research Council, Sesto Fiorentino (Firenze), Italy Department of Mathematics and Computer Science, University of Udine, Udine, Italy

Grapevine named ‘MERLOT KANTHUS’

Abstract: A new and distinct variety of grapevine named ‘MERLOT KANTHUS’, primarily adapted to the growing conditions of the temperate regions, and characterized by its medium vigor; erect growth habit; medium sized, circular shaped leaves; small, conical, medium dense berry clusters with two middle size wings; blue-black berries with slightly firm flesh, none to herbaceous taste and none to very weak flesh coloration; primary use for wine; and resistance to winter temperatures (to −22° C), resistance to downy mildew, and tolerance to powdery mildew