siRNA technology
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This document discusses siRNA (small interfering RNA) technology. It defines siRNA as a class of double stranded RNA molecules typically 20-24 base pairs in length that interfere with gene expression through mRNA degradation. The document outlines the structure and function of siRNA, including its role in the RNAi pathway. Applications of siRNA technology discussed include using it as a research tool to study gene function and as a potential therapeutic agent, for example to target the ApoB gene for hypercholesterolemia or VEGF for age-related macular degeneration. Challenges to clinical use of siRNA are delivery and specificity, but it remains a promising approach for molecular medicine.
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RNA INTERFERENCE: UNRAVELING GENETIC SILENCING
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
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Gene silencing by RNA interference & Antisense therapy
This document discusses two methods of gene silencing: RNA interference (RNAi) and antisense therapy. RNAi involves long double-stranded RNA being cut into small interfering RNAs (siRNAs) that bind to messenger RNA and prevent translation. Antisense therapy uses short antisense DNA oligonucleotides that are complementary to mRNA and form DNA-RNA hybrids that are degraded by RNaseH, blocking translation. Both methods allow targeted inhibition of gene expression for research and potential therapeutic applications.
Micro RNARNA INTERFERENCE AND ITS APPLICATIONS IN CROP IMPROVE...
This document provides an overview of RNA interference (RNAi) and its applications in crop improvement. It discusses the history and discovery of RNAi, the mechanism of RNAi involving initiation by Dicer and effector function of RISC complexes, and methods of transforming plants with RNAi constructs. Applications of RNAi described in the document include modifying traits in rice, maize, barley, cotton, jute, tomato, lathyris, and coffee to improve nutritional quality, increase yields, confer virus resistance, and remove toxic compounds.
Rnases in anti-Viral and anti-Cancer therapy
RNA-based therapies such as mRNA vaccines and ribonucleases are emerging as promising immunotherapies for cancer and infectious diseases. mRNA can be engineered to transiently express proteins and has entered clinical trials for cancer immunotherapies and infectious disease vaccines. Ribonucleases are enzymes that degrade RNA and have cytotoxic effects, making them a potential therapeutic for cancer by selectively targeting and destroying tumor cells. Several RNA-based therapeutics are currently in clinical testing for a range of diseases. mRNA in particular has potential for cost-effective scalable production and for inducing protective immune responses with small doses.
Gene Silencing
This document discusses gene silencing, which is the suppression or interruption of gene expression at the transcriptional or translational level. It regulates gene expression and prevents certain genes from being expressed. There are several types of gene silencing, including transcriptional silencing through genomic imprinting, paramutation, and transposon silencing. Post-transcriptional gene silencing occurs through RNA interference pathways using small interfering RNA, microRNA, Dicer, and RISC complex. Research methods for gene silencing include antisense oligonucleotides, ribozymes, siRNA, and microRNA. Gene silencing has applications in biotechnology and medicine, with advantages such as being cost effective and having high specificity, though it also has disadvantages like potential toxicity.
RNA Interference (RNAi) and RNA Induced Gene Silencing
RNA interference (RNAi) is a natural process in cells that uses small RNA molecules to regulate gene expression. Craig Fire and Andrew Mello discovered RNAi in 1998 through their experiments injecting double-stranded RNA into C. elegans worms. They found this silenced or interfered with the expression of genes with complementary RNA sequences. RNAi has important roles in defending against viruses and regulating development, with the small RNAs binding to messenger RNAs to decrease their activity. It has potential applications in gene therapy, functional genomics, agriculture, and more.
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Antisense RNA technology & its role in crop improvement ppt surendra singh
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Pe gylated drug delivery systems for si rna drug development in cancer therapy
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Antisense rna experiments lead to the discovery of
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Gene silencing by RNA interference & Antisense therapy
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Micro RNARNA INTERFERENCE AND ITS APPLICATIONS IN CROP IMPROVE...
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RNA Interference (RNAi) and RNA Induced Gene Silencing
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Best Ayurvedic medicine for Gas and Indigestion
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Physiology of Special Chemical Sensation of Taste
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
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Top-Vitamin-Supplement-Brands-in-India List
Swisschem Dermacare provides the Top 10 Vitamin Supplement Brands in India. To know more about us give us call at our official number
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- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Novas diretrizes da OMS para os cuidados perinatais de mais qualidade
Novas diretrizes da OMS para os cuidados perinatais de mais qualidadeProf. Marcus Renato de Carvalho
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
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siRNA technology
- 1. siRNA TECHNOLOGY presented by RIA DAS M.Pharm (PHARMACOLOGY) 1st semester, 1st year Registration Number:211002320210008 Roll Number:10020221008 Department Of Pharmaceutical Science And Technology MAULANA ABUL KALAM AZAD UNIVERSITY OF TECHNOLOGY
- 2. CONTENTS: RNA What is RNA interference (RNAi)? siRNA Properties of small non-coding RNA Structure of siRNA RNAi: two phases siRNA pathway illustration Function of siRNA Applications of siRNA siRNA targeting ApoB siRNA for treatment of AMD Advantages of siRNA Challenges Conclusion References
- 3. RNA: RNA or Ribonucleic Acid is a polymer essential in various biological roles in coding, decoding, regulation, and expression of genes. It is made up of Ribose sugar, phosphate, and nitrogenous bases Adenine, Guanine, Cytosine and Uracil.
- 4. What is RNA interference (RNAi)? “The process by which dsRNA silences gene expression” RNAi is the mechanism that inhibits gene expression at a stage of translation or by hindering the transcription of specific genes. Highly specific process. Very potent activity. RNAi targets include RNA from viruses and transposons. Two types of small RNA molecules - miRNA and siRNA are central to RNA interference.
- 5. siRNA: siRNA or Small interfering RNA or sometimes known as short interfering RNA or silencing RNA. Class of double stranded RNA. Non-coding RNA molecules. Typically 20-24 base pairs in length. It interferes with the expression of specific genes with complementary nucleotide sequences by degrading mRNA after transcription, preventing translation.
- 6. Properties of small non-coding RNA: Involved in silencing of other mRNA transcripts. Silences an mRNA by base pairing with some sequence on the mRNA. Usually they are only about 20-24 nucleotides long. Synthesized by first cutting up longer precursor sequences.
- 7. Structure of siRNA: Naturally occurring siRNAs have a well defined structure. It is short, usually of 20-24 base pairs. Double stranded RNA. Phosphorylated 5’ ends, and hydroxylated 3’ ends.
- 8. RNAi: two phases Initiation Generation of mature siRNA. Execution Silencing of target gene. Degradation or inhibition of translation.
- 10. Function of siRNA: Maintain genome integrity against foreign RNA molecules. As therapeutic agent.
- 11. Applications of siRNA: Basic Research: Determining protein function. Clinical Research: Cancer, hypercholesterolemia, infections, developmental defects.
- 12. siRNA targeting ApoB: ApoB, a liver enzyme essential for assembly and secretion of low-density lipoprotein (LDL). It is required for metabolism of cholesterol. High levels of ApoB and LDL increase the risk of coronary artery disease. Cholesterol-conjugated siRNA targets ApoB enzyme and checks the enzyme production. Alnylam Pharmaceuticals developed siRNA to target ApoB and demonstrated the production effect on mice first.
- 13. siRNA for treatment of AMD: Age-related macular degeneration (AMD) is an eye disease that destroys central vision by damaging macula, the central region of retina. Patients may develop a blank or blind spot in their central field of vision. Objects may appear distorted or smaller than they really are. Straight lines may appear wavy or curved. There are no effective therapies. AMD is often associated and prompted by neovascularization. Macular neovascularization is stimulated by interaction of vascular endothelial growth factor (VEGF) with vascular endothelial growth factor receptor (VEGFR-1). Inhibiting production of VEGFR-1 should stop neovascularization and prevent AMD.
- 14. Advantages of siRNA: Cost effective. Highly specific method. Regulation of target gene. Helpful tool for gene function analysis. Helpful in gene therapy.
- 15. Challenges: Not been fully approved in clinical practice. Patient might develop an immune response against the therapy. Target delivery. Off-target effects.
- 16. Conclusion: siRNA have become not only an exciting new tool in molecular biology, but also the next frontier in molecular medicine. Significant hurdles remain, most notably guaranteeing specificity and finding safe and efficacious delivery system. Work is ongoing to solve these problem, but the therapeutic promise of siRNA remains great.
- 17. References: Khvorova, A., Reynolds, A. and Jayasena, S.D. (2003) Functional siRNAs and miRNAs exhibit strand bias. Cell 115, 209–216. Morris, K.V., Chan, S.W., Jacobsen, S.E. and Looney, D.J. (2004) Small interfering RNA-induced transcriptional gene silencing in human cells. Science 305, 1289–1292. Kawasaki, H. and Taira, K. (2004) Induction of DNA methylation and gene silencing by short interfering RNAs in human cells. Nature 431, 211–217. Reynolds, A. et al. (2004) Rational siRNA design for RNA interference. Nat. Biotechnol. 22, 326–330.