Dark Blue and White Biology Education Presentation

RNA-based Therapeutics.

Prepared by:. Rawan Magdy Hamdy-191357 (Introduction ) 4,5 Maryam Ashraf Islam-192055 (Introduction ) 6,7 Fatima Raafat Abdelhady-192129 (Principle ) 8,9 Nehal Khaled Gadelkareem-194237 (Principle ) 10,11 Aliaa Akram Mohamed-191435 (Principle ) 12,13,14,15 Klodia Hani-190151 (Applications ) 16,17 Kerolos Hazem Habashy-192045 (Applications ) 18,19,20,21.

Outline:. Background introduction containing definition and types. Principle of the previous technique. Applications RNA-based therapeutics ..

Introduction:. Numerous discoveries have established RNA therapy as an indispensable technology for treating human disease. RNA was first described as a key player in the flow of genetic information by Crick in his study “Central Dogma of Molecular Biology” and was later confirmed by the discovery of mRNA, which highlighted the importance of these molecules as key messengers in the translation of.

Introduction:. The use of RNA-based compounds to modify biological pathways in order to treat a particular ailment is known as "RNA therapy." Generally speaking, controlling the expression or activity of a target molecule requires.

Introduction:. According to Fire (1998), Once the nucleic acid chemistry and the delivery method are established, the production of RNA-based drugs for a new target can be achieved in a relatively short period using these pre-established methodologies. It is this strength that has made messenger RNA (mRNA) therapy, a subcategory of RNA therapy, the most important and efficient means for developing novel.

siRNA miRNA so RNA-based Therapeutics Ribozyme Aptamer.

RNA Interference ( RNAi ):. Double-stranded RNA-mediated interference ( RNAi ) is a simple and rapid method of silencing gene expression in a range of organisms that cause disease or that contribute to disease. Extensive genetic and biochemical analysis revealed a two-step mechanism of RNAi -induced gene silencing. The first step ( RNAi initiating step) involves degradation of dsRNA into small interfering RNAs ( siRNAs ) In the second step, the siRNAs join an RNase complex, RISC (RNA-induced silencing complex), which acts on the cognate mRNA and degrades it..

RNA Interference ( RNAi ):. RNA interference serves a role in protecting cells from viruses, by attacking their mRNAs and sometimes even their RNA-based genomes before the cell’s synthesis machinery can manufacture viral proteins. By targeting mRNAs rather than proteins, RNA interference stands to work around the resistance that can complicate treatment for many conditions. This can make medicine for even complex situations more specific while reducing side effects (Adams, 2018)..

Antisense oligonucleotides (ASOs):. What is antisense oligonucleotide?.

Antisense oligonucleotides (ASOs):. Binds to RNA and hinder its function without facilitating RNA degradation such as translation inhibition or modulation of RNA processing. Promotes degradation of RNA via: endogenous enzymes such as RNase H or argonaute-2 (RNA interface (RNAI). Also can increase protein production by binding to sequences in upstream of primary ORF or by antagonizing miRNAs , which normally diminish protein production (Yang, 2018)..

CRISPR/CAS -based RNA editing system:. Principle.

CRISPR/CAS 9-based RNA editing system:. Principle.

CRISPR/CAS-based RNA editing systems:. Principle.

CRISPR/CAS 13-based RNA editing system:. Principle.

Dva. Applications:. CRISPR/Cas-based genome therapy: The first human trial using CRISPR/Cas to treat genetic diseases was β-thalassemia (NCT03655678). Also, they used this method to treat retinal defect by EDIT-101 (NCT03872479) (Mullard,2017)..

Applications:. mRNA vaccine: According to Doudna (2021), They are made to treat virus infection such as corona virus as they made the Pfizer- BioNTech vaccine BNT162b2 ( Comirnaty ®) and the Moderna vaccine mRNA-1273 to treat the disease and they were the first two FDA-approved SARS-CoV-2 vaccines, which both proven >94% efficacy in phase III clinical trials..

Applications:. Disease disorder: O Mutation at exon 51 of dystrophin mRNA. O Inducing premature translation termination. O Therefore, prevent production of functional protein..

Applications:. Drug mechanism: O It binds to “ exonic splicing enhancer”. O Induces “exon 51” skipping. O Therefore, reduces percentage of produces fully functional proteins..

a Mechanism of action for ASO .ASQ. RNaseH Splicing RNA degradation b Mipomersen Translation ApoB•100 protein ApoB.100 mRNA + Mipomersen RNase H mRNA degradation.

C Nusinersen SMNI pre-mRNA Normal splicing Exon 7 inclusion Translation Stable SMN Nusinersen SMN2 pre-mRNA Nusinersen 5 6 Intronic splicing silencer Normal splicing Exon 7 8 skipping Translation d Eteplirsen DMO pre-mRNA of patient Eteplirsen 4 VVithout treatment pre-mature termination No protein Unstable SMN Exonic splicing enhancer + Eteplirsen 4 Normal reading frame Shortened dystrophin.

References:. Crick, FH On protein synthesis. Symp Soc Exp Biol 12, 138–163 (1958). Brenner, S, Jacob, F & Meselson , M An unstable intermediate carrying information from genes to ribosomes for protein synthesis. Nature 190, 576–581 (1961). Gros , F et al. Unstable ribonucleic acid revealed by pulse labelling of Escherichia coli. Nature 190, 581–585 (1961). Fire, A et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans . Nature 391, 806–811 (1998). Adams, D et al. Patisiran , an RNAi therapeutic, for hereditary transthyretin amyloidosis. N Engl J Med 379, 11–21 (2018). Reich, E, Franklin, RM, Shatkin , AJ & Tatumel Action of actinomycin D on animal cells and viruses. Proc Natl Acad Sci USA 48, 1238–1245 (1962). Mullard A. Small molecules against RNA targets attract big backers. Nat Rev Drug Discov . 2017;16:813–5. Doudna JA, Charpentier E. Genome editing. The new frontier of genome engineering with CRISPR-Cas9. Science. 2021;346:1258096. Asmamaw, M. and Zawdie , B. (2021). Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing. Biologics : Targets & Therapy, [online] 15, pp.353–361. doi:10.2147/BTT.S326422. Li, X.; Yang, L.; Chen, L. L. The Biogenesis, Functions, and Challenges of Circular RNAs. Mol. Cell 2018, 71, 428– 442, DOI: 10.1016/j.molcel.2018.06.034 ..

Thanks! Any questions?.