Metodi post-genomici in biochimica cellulare. Metodi post-genomici.

Presentazione sul tema: "Metodi post-genomici in biochimica cellulare. Metodi post-genomici."— Transcript della presentazione:

1 Metodi post-genomici in biochimica cellulare

2 Metodi post-genomici

3 Quantitative analysis of systems biology by taking advantage of available genomic information at the level of 1.SNPs analysis associated to disease or drug response 2.mRNA (transcriptomics) 3.Protein (proteomics) 4.Post-translational modifications (aka “modificomics”) 5.Surface exposure (surfomics) 6.Protein-protein interactions (interactomics) n 7.Small metabolites (metabolomics) and their relations (metabonomics) Many other fantasy exercises (glycomics, lipidomics, allergenomics, degradomics, excluding – perhaps – comics...) G.B Smejkal, “I’m an –omics, you’re an -omics... ” Exp. Rev. Proteomics 3 (2006) 383-385 Metodi post-genomici

4 Modelli cellulari e animali Trascrittomica Proteomica Systems biology

5 Modelli cellulari e animali (farmacologici e/o genetici)

6 Modelli cellulari (farmacologici e/o genetici) Facilità di mantenimento e trattamento Possibilità di combinare trattamento farmacologico e manipolazione genetica Utili per riprodurre un singolo meccanismo Cellule umane (o murine)

7 Modelli animali (farmacologici) Intero organismo vs. cellule isolate Trattamento sistemico o lesione chimica locale Possibilità di valutare l’effetto anterogrado/retrogrado

8 Modelli animali (e vegetali?!?) (genetici) Organismi modello Genoma noto Non solo topo! Vita breve Invertebrati (e piante…)

9 Trascrittomica

10 Common steps to expression analysis

11 RTqPCR microRNAs are short (around 23 nt) non coding RNAs with roles in transcriptional regulation of gene expression

12 RTqPCR

13

14 Quantification & Analysis

15

16 Arrays

17

18 Microarray technology has two major applications: gene expression analysis and genetic variation analysis. All gene expression microarrays are based on probes hybridization of labelled cDNA. Differences arise in the labelling dye, deployment, and probes immobilization and/or organization. Illumina beads arrays Affymetrix genechips Agilent microarrays

19

20 Cluster analysis

21 Euclidean distance Pearson correlation

22 Do it yourself:

23 Next generation sequencing

24 1 st gen. Sanger seq.

25 2 nd gen. Illumina https://www.youtube.com/watch?v=womKfikWlxM

26 2 nd gen. IonTorrent https://www.youtube.com/watch?v=WYBzbxIfuKs

27 3 rd gen. Pacific Biosciences https://www.youtube.com/watch?v=v8p4ph2MAvI

28 3 rd gen. Oxford Nanopore https://www.youtube.com/watch?v=3UHw22hBpAk