Universitą degli studi di Pavia
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Brandi's curriculum
Curriculum
PERSONAL INFORMATION:
Brandi Ornella, E-mail: ornella.brandi01@universitadipavia.it, Date of birth 16/04/1989 Nationality: Italian.
EDUCATION AND TRAINING:
English: Listening: B2, Reading: C1, Spoken Interaction: B1, Spoken production: B1, Writing: B1. (Levels: A1/A2: Basic user - B1/B2: Independent user - C1/C2: Proficient user Common European Framework of Reference for Languages).
Job-related skills: excellent knowledge of many molecular biology techniques : electrophoresis on agarose gel, plasmid cloning, PCR, plasmid DNA purification, DNA gel extraction, genomic and plasmid DNAextraction, qualitative and Real Time PCR, RNA and protein extraction, retrotranscription of RNA in cDNA, Bradford assay, cell transfection, DNA sequencing, SDS-page and western blot analysis, transcription reactions, in situ hybridization, immunofluorescence; cell cultures; Bioinformatics : "Deep view" (protein structure analysis software); "Serial cloner" (plasmid cloning software); nucleotide and protein sequences alignment; use of many databases). Drosophila melanogaster (UAS-Gal4 system, matching schemes, immunofluorescent staining of Drosophila larval tissues, immunofluorescent staining of Drosophila embryos; mitotic recombination; Flip-out system); Skills mainly gained during the academic and post-academic internships.
Computer skills: excellent knowledge of Microsoft Office (Word, Excel, power point); good knowledge of software to create scentific bibliography (EndNote); basic use of Adobe Photoshop CS5.
REFERENCES:
Professor Alessandro Riccardo, Dipartimento di Biopatologia e Biotecnologie Mediche e Forensi, Sezione di Biologia e Genetica, Universitą di Palermo, riccardo.alessandro@unipa.it. Dott.ssa Raimondo Stefania; Dip. DiBiMEF, sez. di Biologia e Genetica; stefania.raimondo@unipa.it. Dott. Bernardoni Roberto, Research associate; Alma mater studiorum, universitą di Bologna; roberto.bernardoni@unibo.it. Dott. Reale Stefano, leader of the laboratory "Tecnologie Diagnostiche Innovative", Istituto Zooprofilattico Sperimentale della Sicilia; stefano.reale@izssicilia.it. Dott. Bellia Chiara, researcher at the "Dipartimento di Biopatologia e Biotecnologie Mediche e Forensi"; chiara.bellia@unipa.it
Pubblicazioni e comunicazioni a congressi
Research project
Use of DNA-damage induced RNA (DDRNA) mimics to enforce DNA damage response activation in the absence of DNA lesions
Our group and others recently published that the safeguard of DNA integrity depends on a novel class of short non-coding RNAs called DNA damage response RNAs (DDRNAs) that act at DNA lesions to promotea DNA damage response (DDR)(FranciaS. et al. Nature 2012; Wei Wei et al Cell 2012; Michalick et al NAR 2012).The role of DDRNAs in DDR was establishedin a conveniently engineered cellularsystem,which allows the controlled induction of a single DNA lesion in a defined and traceable locus. With this cell system,we showed thatchemically synthesized DDRNAs homologous in sequence to DNA flanking the damage site, control DDR fociformation in a dose dependent manner. More recently, we obtained unpublished data demonstrating that DDRNAs act at sites of DNA damage by pairing to nascent transcripts emerging from DNA Double strand breaks (DSBs) (Michelini F. et al submitted)and promote the accumulation of DDR mediators to site of DNA damagebutare dispensable for the initial recognition of DNA lesion (Francia S. et al. Submitted). Since the tethering of key DDR factors to an genetic locus is sufficient to trigger full DDR signaling evenin the absence of physical DNA lesions(Soutoglou E., Science 2008),we hypothesized that exogenously introduced DDRNAs may be sufficient to stimulate DDR factors recruitment and DDR activation in target cells. Indeed, preliminary datafrom our group show thattransfection of synthetic DDRNAs induces DDR activation on acomplementary genomic locus, and this occursin a relevant fraction of cellseven in the absence of a preexisting DNA lesion. Prompted by these preliminary results, we want to test if the formation of a DDR focus induced by DDRNAs transfection in the absence of DNA lesions, also leads to the activation of a DNA damage cell-cycle checkpoint specifically in cells bearing target sequences, with the consequence of reducing their proliferation. This could give a selective proliferative disadvantage to cells harboring a specific genetic mutation in a mixed population, with potential relevant implications. To test this intriguing hypothesis, we will usea mixed population oftarget and parental cell lines, whichonly differ for the presence of an engineeredtarget genetic locusand we will transfect it with sequence-specific DDRNAs,or sequence-unrelated oligos as control.Upon multiple rounds of transfection, we will measure the clonal expansion of the two cell lines in time. Measurements will be performed by Q-PCR on genomic DNA, using cell type specific primers. Results obtained will be confirmed in the different cellular systems (NIH3T3,HCT 116 and HT1080)and, if the hypothesis holds true in in vitro experiments, we aim to confirm it by an in vivo approach, targeting with DDRNAS conveniently chosen cancer cell lines inside tumors formed in nude mice.
In summary, with this projectwe will testthe possibility that sequence-specific, synthetic DDRNAsmay be usedto induce a selective proliferative disadvantage to a specific target cell population bearing genetic alterations such as mutations orviral integrations, even in the absence of physical DNA damage.
PERSONAL INFORMATION:
Brandi Ornella, E-mail: ornella.brandi01@universitadipavia.it, Date of birth 16/04/1989 Nationality: Italian.
EDUCATION AND TRAINING:
- 02/11/2015–present: Phd student in "Genetics, molecular and cell biology", Universitą degli studi di Pavia; Research activities at the Istituto di Genetica Molecolare (IGM-CNR) of Pavia.
- 10/2015: Gain of a position for the Phd program in "Experimental Oncology and surgery" (international). Position rejected. Universitą degli studi di Palermo.
- 04/2015–08/2015: Internship as a volunteer at a molecular and cell biology laboratory. Dip. DiBiMEF, Biology and Genetics, Via Divisi 83, Palermo (Italia).
- 12/2014: Qualified as a biologist. Universitą degli studi di Parma.
- 10/2014: Gain of a scholarship for the Phd program in “ Life and Health applied Sciences”. Rejected. Universitą degli studi di Verona. Position was rejected because of the poor interest toward the related research project.
- 01/2012–03/2014: Master's degree, Molecular and Industrial Biotechnology. Alma Mater studiorum-universitą di Bologna. Date: 20/03/2014; Vote: 110/110 with honors.
- 03/2013–03/2014: Academic Internship (genetics). Alma Mater studiorum-universitą di Bologna, Bologna (Italia). Department of pharmacy and biotechnology. Genetics laboratory (Drosophila melanogaster), Dott. Bernardoni Roberto, via Selmi n°3. Thesis: "In vivo study of the function of the Multidrug-Resistance likeProtein 1 gene, in Drosophila melanogaster (dMRP)".
- 10/2008–12/2011: Bachelor’s Degree in Biotechnology. Universitą degli studi di Palermo, Palermo (Italy). Date: 14/12/2011; Vote: 110/110 with honors.
- 09/2011–12/2011: Academic Internship (microbiology). Istituto Zooprofilattico Sperimentale della Sicilia ("A. Mirri"), Palermo (Italy).Molecular biology, Dott. Reale Stefano. Thesis: "Use of the Micro-Seq technique for the identification of microbial and fungal species on biological matrices "
English: Listening: B2, Reading: C1, Spoken Interaction: B1, Spoken production: B1, Writing: B1. (Levels: A1/A2: Basic user - B1/B2: Independent user - C1/C2: Proficient user Common European Framework of Reference for Languages).
Job-related skills: excellent knowledge of many molecular biology techniques : electrophoresis on agarose gel, plasmid cloning, PCR, plasmid DNA purification, DNA gel extraction, genomic and plasmid DNAextraction, qualitative and Real Time PCR, RNA and protein extraction, retrotranscription of RNA in cDNA, Bradford assay, cell transfection, DNA sequencing, SDS-page and western blot analysis, transcription reactions, in situ hybridization, immunofluorescence; cell cultures; Bioinformatics : "Deep view" (protein structure analysis software); "Serial cloner" (plasmid cloning software); nucleotide and protein sequences alignment; use of many databases). Drosophila melanogaster (UAS-Gal4 system, matching schemes, immunofluorescent staining of Drosophila larval tissues, immunofluorescent staining of Drosophila embryos; mitotic recombination; Flip-out system); Skills mainly gained during the academic and post-academic internships.
Computer skills: excellent knowledge of Microsoft Office (Word, Excel, power point); good knowledge of software to create scentific bibliography (EndNote); basic use of Adobe Photoshop CS5.
REFERENCES:
Professor Alessandro Riccardo, Dipartimento di Biopatologia e Biotecnologie Mediche e Forensi, Sezione di Biologia e Genetica, Universitą di Palermo, riccardo.alessandro@unipa.it. Dott.ssa Raimondo Stefania; Dip. DiBiMEF, sez. di Biologia e Genetica; stefania.raimondo@unipa.it. Dott. Bernardoni Roberto, Research associate; Alma mater studiorum, universitą di Bologna; roberto.bernardoni@unibo.it. Dott. Reale Stefano, leader of the laboratory "Tecnologie Diagnostiche Innovative", Istituto Zooprofilattico Sperimentale della Sicilia; stefano.reale@izssicilia.it. Dott. Bellia Chiara, researcher at the "Dipartimento di Biopatologia e Biotecnologie Mediche e Forensi"; chiara.bellia@unipa.it
Pubblicazioni e comunicazioni a congressi
Research project
Use of DNA-damage induced RNA (DDRNA) mimics to enforce DNA damage response activation in the absence of DNA lesions
Our group and others recently published that the safeguard of DNA integrity depends on a novel class of short non-coding RNAs called DNA damage response RNAs (DDRNAs) that act at DNA lesions to promotea DNA damage response (DDR)(FranciaS. et al. Nature 2012; Wei Wei et al Cell 2012; Michalick et al NAR 2012).The role of DDRNAs in DDR was establishedin a conveniently engineered cellularsystem,which allows the controlled induction of a single DNA lesion in a defined and traceable locus. With this cell system,we showed thatchemically synthesized DDRNAs homologous in sequence to DNA flanking the damage site, control DDR fociformation in a dose dependent manner. More recently, we obtained unpublished data demonstrating that DDRNAs act at sites of DNA damage by pairing to nascent transcripts emerging from DNA Double strand breaks (DSBs) (Michelini F. et al submitted)and promote the accumulation of DDR mediators to site of DNA damagebutare dispensable for the initial recognition of DNA lesion (Francia S. et al. Submitted). Since the tethering of key DDR factors to an genetic locus is sufficient to trigger full DDR signaling evenin the absence of physical DNA lesions(Soutoglou E., Science 2008),we hypothesized that exogenously introduced DDRNAs may be sufficient to stimulate DDR factors recruitment and DDR activation in target cells. Indeed, preliminary datafrom our group show thattransfection of synthetic DDRNAs induces DDR activation on acomplementary genomic locus, and this occursin a relevant fraction of cellseven in the absence of a preexisting DNA lesion. Prompted by these preliminary results, we want to test if the formation of a DDR focus induced by DDRNAs transfection in the absence of DNA lesions, also leads to the activation of a DNA damage cell-cycle checkpoint specifically in cells bearing target sequences, with the consequence of reducing their proliferation. This could give a selective proliferative disadvantage to cells harboring a specific genetic mutation in a mixed population, with potential relevant implications. To test this intriguing hypothesis, we will usea mixed population oftarget and parental cell lines, whichonly differ for the presence of an engineeredtarget genetic locusand we will transfect it with sequence-specific DDRNAs,or sequence-unrelated oligos as control.Upon multiple rounds of transfection, we will measure the clonal expansion of the two cell lines in time. Measurements will be performed by Q-PCR on genomic DNA, using cell type specific primers. Results obtained will be confirmed in the different cellular systems (NIH3T3,HCT 116 and HT1080)and, if the hypothesis holds true in in vitro experiments, we aim to confirm it by an in vivo approach, targeting with DDRNAS conveniently chosen cancer cell lines inside tumors formed in nude mice.
In summary, with this projectwe will testthe possibility that sequence-specific, synthetic DDRNAsmay be usedto induce a selective proliferative disadvantage to a specific target cell population bearing genetic alterations such as mutations orviral integrations, even in the absence of physical DNA damage.
