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Biochimica II
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Anno accademico 2010/2011
- Codice dell'attività didattica
- B8030
- Docente
- Dott. Riccardo Taulli (Titolare del corso)
- Corso di studi
- laurea i^ liv. in biotecnologie - a torino
- Anno
- 3° anno
- Tipologia
- Bio-Molecolare
- Crediti/Valenza
- 5
- SSD dell'attività didattica
- BIO/10 - biochimica
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Sommario insegnamento
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Obiettivi formativi
Regolazione post-trascrizionale dell'espressione genica. Il corso è mirato all'apprendimento dei 1)meccanismi biochimici alla base della maturazione dei piccoli RNA non codificanti: siRNAs e microRNAs 2)meccanismi di regolazione genica a livello post-trascrizionale 3) le libreria ad siRNAs e i vettori lentivirali come nuovi ed efficaci strumenti per lo studio della funzione genica in vitro ed in vivo 4) il ruolo dei microRNAs nella regolazione dei processi fisiologici (sviluppo muscolare e cardiaco, l'ematopoiesi) 5) il ruolo dei microRNAs nel cancro
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Risultati dell'apprendimento attesi
Alla fine del corso lo studente avrà acquisito le conoscenze dei meccanismi di regolazione della funzione genica a livello post-trascrizionale mediata sia da siRNAs che microRNAs. Inoltre, lo studente avrà acquisito le competenze per un uso corretto in laboratorio della tecnologia del silenziamento genico post-trascrizionale. Infine, lo studente avrà aquisito le competenze per comprendere il ruolo dei microRNAs nei processi fisiologici e nella trasformazione tumorale.
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Programma
Modulo I: RNA interference
· RNAi mechanism, microRNA pathway
· similarities and differences between miRNA and siRNA pathway and mechanism
· the Cosuppression mechanism in plants
· RNAi transitive in Plants and C. Elegans: the role of Sid-1 protein and RpRP in RNAi transitive and in diffusion process
· RNAi in mammalian cells
- siRNA advantage and disavantage
- shRNA advantage and disavantage
- systems with high efficency of transduction: retrovirus and lentivirus
- lentiviral vector for conditional expression of shRNA:
a) Cre-Lox system
b) Tet-On/Tet Off system (based on Pol III promoter)
c) Tet-On/Tet Off system (based on Pol II proter and miR-30 based backbone)
- RNAi libraries:
- first generation of shRNAs library (Ex Hannon GJ. and Bernards R. shRNAs-based library)
- synthetic shRNAs as potent RNAi trigger
- second generation of shRNAs library (Hannon G.J. shRNAs-based library).
- RNAi and Off-target effects
- Application of RNAi techonology
analysis of gene function using RNAi library, identification of new pharmacological targets
siRNAs as possible future innovative drugs (presentation of RIGHT project)
in vivo optical imaging and MRI of siRNAs
RNAi transgenesis as a tool to study gene function in vivo rapidly and at low cost
Modulo II: microRNA biogenesis and functions
· microRNAs story: Lin-4 in C. elegans.
· Identification of miRNAs in mammalian (from mouse to human)
· nomenclature and miRNAs registry
· distribution of miRNAs in the genome
expression of microRNAs
microRNAs biogenesis and mechanism
1. microRNAs supress mRNA translation
2. microRNAs promote mRNA degradation (miRNAs as siRNAs)
3. microRNAs direct mRNA deadenylation
4. microRNA promote gene expression
· biological function of miRNAs in mammalian
- miRNAs function in early vertebrate development
- miRNAs function in late vertebrate development
· the impact of mammalian miRNAs on mRNA repression and evolution
· microRNA and Cancer
- microRNA and fragile sites
- classification of human cancer by microRNAs
- post-trascriptional regulation of the let-7 microRNA during neural cell specification
- extensive post-trascriptional regulation of microRNA and its implication in cancer
- microRNAs as oncosuppressors:
a) the role of miR-15 and Mir-16 in CLL, regulation of BCL-2 by miR-15/16
b) the role of Let-7 in lung cancer, regulation of Ras by Let-7 familyù
c) Let-7 supresses tumorigenesis Ras-dependent in vivo
d) a microRNA component of the p53 tumor suppressor netwok
e) endogenous microRNAs that supress brest cancer metastasis
microRNAs as oncogenes:
a) the role of polycistronic microRNA cluster in lung cancer
b) the polycistronic microRNA as a potential human oncogene
c) the oncogenic role of mir 372/373
- microRNAs as indicators of tumor progression and prognosis
- microRNA in heart and muscle development
a) the role of miR-1 and miR 133 in muscle proliferation and differentiation
b) Muscle-specifi c microRNA miR-206 promotes muscle differentiation
c) the physio-pathological functions of miR-1and miR-133 in the heart
d) MicroRNA-133 controls cardiac hypertrophy
e) Dysregulation of Cardiogenesis, Cardiac Conduction, and Cell Cycle in Mice Lacking miRNA-1-2
f) Control of stress-dependent cardiac growth and gene expression by a microRNA.
- microRNAs in hematopoietic differentiation
a) the role of miR-181 in B-cell lineage
b) the role of miR-221 and miR-222 in erithropoiesis
c) the role of miR-223 in APL differentiation
d) Regulation of progenitor cell proliferation and granulocyte function by microRNA-223.
- microRNA and tissue specific “gene therapy”
microRNA as possible future innovative drugs or pharmacological target:
a) the Antagomir
microRNA and imprinting
Dicer ablation affects B lynphocyte Lineage
Targeted deletion reveals essential and overlapping functions of the miR-17 through 92 family of miRNA clusters.
Lymphoproliferative disease and autoimmunity in mice with increased miR-17-92 expression in lymphocytes.
Testi consigliati e bibliografia
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Articoli scientifici e materiale didattico fornito dal Docente
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Note
Modalità di Esame: Colloquio Orale
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