1. Regulatory Relationship of microRNAs in Alzheimer and Huntington Diseases: A Cladistics Approach Prachi Srivastava AMITY Institute of Biotechnology AMITY University Uttar Pradesh Lucknow Lucknow, UP, India

2. AMITY INSTITUTE OF BIOTECHNOLOGY INTRODUCTION 2

3. AMITY INSTITUTE OF BIOTECHNOLOGY RNA interference (RNAi) RNA interference (RNAi) is a highly evolutionally conserved process of post-transcriptional gene silencing (PTGS) double stranded RNA (dsRNA), when introduced into a cell, causes sequence-specific degradation of homogolous mRNA sequences. It was first discovered in 1998 by Andrew Fire and Craig Mello in the nematode worm Caenorhabditis elegans and later found in a wide variety of organisms, including mammals

4. AMITY INSTITUTE OF BIOTECHNOLOGY T IMELINE 1998 1999 2000 2001 2002 1990 cosuppression of purple color in plants dsRNA injection in worms short RNAs identified in plants RNAi shown in vitro RISC activity partially purified siRNAs identifiedDicer identified RNAi used against HIV genome-wide RNAi screens begin

5. AMITY INSTITUTE OF BIOTECHNOLOGY RNA Interference Phenomena first observed in petunia Attempted to overexpress chalone synthase (anthrocyanin pigment gene) in petunia. (trying to darken flower color) Caused the loss of pigment. Called co-suppression because suppressed expression of both endogenous gene and transgene

6. AMITY INSTITUTE OF BIOTECHNOLOGY RNA Interference Approaches Four types of responses induced by dsRNA 6

7. AMITY INSTITUTE OF BIOTECHNOLOGY The Mechanism of RNAi 7

8. AMITY INSTITUTE OF BIOTECHNOLOGY MicroRNA(miRNA) 8

9. AMITY INSTITUTE OF BIOTECHNOLOGY miRNA vs. siRNA 9

10. AMITY INSTITUTE OF BIOTECHNOLOGY Integration of RNAi in Drug Discovery 10

11. AMITY INSTITUTE OF BIOTECHNOLOGY RNAi based therapeutics 11

12. AMITY INSTITUTE OF BIOTECHNOLOGY RNA-interference-based therapies 12

13. AMITY INSTITUTE OF BIOTECHNOLOGY 13

14. AMITY INSTITUTE OF BIOTECHNOLOGY METHODOLOGY 14

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20. AMITY INSTITUTE OF BIOTECHNOLOGY RESULT AND CONCLUSION 20

21. AMITY INSTITUTE OF BIOTECHNOLOGY Alzheimer and Huntington 21

22. AMITY INSTITUTE OF BIOTECHNOLOGY Alzheimer and Huntington S no.miRNAGene familyRegulation patternBootstrap score 1.Mir-22(H)MIPF0000053Down Regulated10 Mir-22(A) 2.Mir-330(H)MIPF0000200Up Regulated5.17 Mir-34a(A)MIPF0000039Down Regulated 3.Mir-132(H)MIPF0000065Down Regulated5.00 Mir-298(A)MIPF0000206Down Regulated 4.Mir-128-1(H)MIPF0000065Down Regulated10 Mir-128-1(A)Up Regulated 5.Mir-29a(H)MIPF0000009Up Regulated10 Mir-29a(A)Down Regulated 22

23. AMITY INSTITUTE OF BIOTECHNOLOGY Alzheimer and Parkinson 23

24. AMITY INSTITUTE OF BIOTECHNOLOGY Alzheimer and Parkinson S no.miRNAGene family Regulation pattern Bootstrap score 1.Mir-26a-1(P)MIPF0000043Down regulated10 Mir-26b(A) 2.Mir-184(P)MIPF0000059N/A- Mir-320(A)MIPF0000163Up regulated 24

25. AMITY INSTITUTE OF BIOTECHNOLOGY Parkinson and Huntington 25

26. AMITY INSTITUTE OF BIOTECHNOLOGY Parkinson and Huntington S no.miRNAGene familyRegulation patternBootstrap score 1.Mir-184(P)MIPF0000059Down regulated10 Mir-132(H)MIPF0000065 2.Mir-133(P)MIPF0000029Down regulated6 Mir-128-1(H)MIPF0000048 26

27. AMITY INSTITUTE OF BIOTECHNOLOGY CONCLUSION Phylogenetic analysis of miRNA of Alzheimer and Huntington diseases gives insight into evolutionary relationship and reveals regulation of Mir-22, Mir 29a, and mir-128-1 in both diseases. Mir-22 shows down-regulation in Parkinson disease and Alzheimer disease while mir-128-1 and mir-29a shows the difference in their regulation pattern In Alzheimer disease mir-128-1 is up regulated while in Huntington disease it is down regulated. Mir-29a is up regulated in Huntington disease while in Alzheimer disease, it is down regulated. These findings illustrates the importance of miRNA research in Neurodegenerative diseases with reference to novel targets identification which can give a better lead in concern to protective or prophylective approaches. 27

28. AMITY INSTITUTE OF BIOTECHNOLOGY REFERENCES 1.Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and Function. Cell 2004;116:281–97. Gregory RI, Chendrimada TP, Cooch N, Shiekhattar R. Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. Cell 2005;123:631–40. 2.Perron MP, Provost P. Protein components of the microRNA pathway and human diseases. Methods Mol Biol 2009;487:369–85. 3.Griffiths-Jones S, Grocock RJ, van Dongen S, Bateman A, Enright AJ. MiRBase: microRNA sequences, targets and gene nomenclature. Nucleic Acids Res 2005;34:D140–4. 4. Luciano DJ, Mirsky H, Vendetti NJ, Maas S. RNA editing of a miRNA precursor. RNA 2004;10:1174–7. John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS. Human microRNA targets. PLoS Biol 2004;2:e363. 5. Lim LP, Lau NC, Garrett-Engele P, Grimson A, Schelter JM, Castle J, et al. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature 2005;433:769–73. 6.Griffiths-Jones S, Saini HK, Van Dongen S, Enright AJ: miRBase: tools for microRNA genomics. Nucleic Acids Res 2008, 36:D154–8. 7.Olena AF, Patton JG: Genomic organization of microRNAs. Journal of cellular physiology 2009, 222:540–545. 8.Altuvia Y, Landgraf P, Lithwick G, Elefant N, Pfeffer S, Aravin A, Brownstein MJ, Tuschl T, Margalit H: Clustering and conservation patterns of human microRNAs. Nucleic Acids Res 2005, 33:2697–2706 9.Kozomara A, Griffiths-Jones S: miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res 2011, 39:D152–7. 10. Felsenstein J: PHYLIP (phylogeny inference package), version 3.5 c. Distributed by the author 1993. 11. Felsenstein J: Parsimony in systematics: biological and statistical issues. Annual review of ecology and systematics 1983, 14:313–333. 12. Zhang R, Wang Y-Q, Su B: Molecular evolution of a primate-specific microRNA family. Mol Biol Evol 2008, 25:1493–1502. 13. Altuvia, Y, Landgraf, P, Lithwick, G, Elefant, N, Pfeffer, S, Aravin, A et al. (2005). Clustering and conservation patterns of human microRNAs. Nucleic Acids Res 33: 2697–2706. 28

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