Lentivirus protocol
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Lentivirus can infect non-dividing cells because its pre-integration complex enters the nucleus and integrates into the host genome. This allows lentivirus to be used as a tool for introducing genes into mammalian and other animal cells. Recombinant lentivirus is generated by cotransfecting 293T cells with an envelope plasmid, packaging plasmid, and lentivirus plasmid containing the gene of interest. The process involves obtaining the target gene by PCR, selecting a lentivirus vector plasmid, constructing the recombinant lentivirus plasmid, and identifying the inserted gene sequence.
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RETROVIRUS MEDIATED GENE TRANSFER AND EXPRESSION CLONING
- The retroviral virion is a spherical particle 80-100 nm in diameter composed of a lipid bilayer envelope containing glycoproteins and a capsid containing two copies of the viral RNA genome and enzymes.
- Retroviruses replicate by reverse transcribing their RNA genome into DNA which is then integrated into the host cell genome by an integrase enzyme to become a provirus, allowing transcription of viral genes.
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Retroviral Vectors
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2. Once inside the cell, the viral genome is released from its protein coat through uncoating.
3. The viral genome then hijacks the host cell machinery to replicate, transcribe mRNA, and translate proteins.
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2. Lentiviruses should be stored at -80°C for long-term storage and diluted in ice water or medium kept at 4°C.
3. Recombinant lentiviruses are produced by co-transfecting lentiviral, envelope, and packaging plasmids into 293T cells, collecting the supernatant, and purifying and titrating the virus.
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Transgenic technology involves introducing exogenous DNA into the genome of an organism. This DNA is then transmitted to progeny. There are several methods for creating transgenic organisms, including embryonic stem cell and pronuclear microinjection methods. The embryonic stem cell method involves inserting foreign DNA into embryonic stem cells, which are then fused with blastula cells. The pronuclear microinjection method injects DNA directly into the male pronucleus of a fertilized egg. Applications of transgenic technology include disease research, agriculture, and pharmaceutical development.
Transfection
The document discusses various methods of transfection in animals. Transfection is the process of introducing nucleic acids into eukaryotic cells. It describes viral transfection using bacteria like Agrobacterium tumefaciens and viruses. Non-viral methods include chemical transfection using calcium phosphate, liposomes, polyamines. Mechanical transfection employs microinjection or particle bombardment. Common chemical methods are calcium phosphate precipitation, polyplexes, and liposomes/lipoplexes. Viruses used are retroviruses, adenoviruses, adeno-associated viruses. Bacterial and viral vectors allow for integration into the host genome while chemical and mechanical are often transient.
Replication-of-Virus.pdf
Viruses replicate inside host cells using the cell's machinery. There are three phases to viral replication: initiation, replication of the viral genome, and assembly/release. Initiation involves attachment and penetration of the host cell. Replication varies depending on whether the virus has DNA or RNA as its genome. RNA viruses must first transcribe their genome into mRNA before it can be translated. DNA viruses can use the host's transcription machinery directly. Assembly involves packaging of new viral particles, which are then released to infect new host cells.
Exprssion vector
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
Cloning vectors
cloning vectors-
pBR322,
pET21,
pUC 18,
COSMID
SV 40
Lambda phage, M13 Bacteriophage, bacterial artificial chromosome,
yeast artificial chromosome, shuttle vector ,expression vector
Animal viral based vectors
Viral vectors are efficient tools for gene delivery due to viruses' ability to transfer DNA into host cells. The document discusses several types of viral vectors, including adenoviral, adeno-associated, retroviral, lentiviral, and baculovirus vectors. It provides details on the structure and genome organization of different viruses used to create these vectors. The document also explains the process of generating recombinant viral vectors by removing unnecessary viral genes and inserting genes of interest. Viral vectors allow for transient or stable gene expression and are useful for both research and clinical applications such as gene therapy and vaccine development.
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Transgenic Animals: The ability to manipulate the genome of the whole animal ...
Transgenic Animals: The ability to manipulate the genome of the whole animal ...
GeneMedi-Lentivirus production protocol-packaging concentration and purification
GeneMedi-Lentivirus production protocol-packaging concentration and purification
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Storyboard on Acne-Innovative Learning-M. pharm. (2nd sem.) Cosmetics
Acne is a common skin condition that occurs when hair follicles become clogged with oil and dead skin cells. It typically manifests as pimples, blackheads, or whiteheads, often on the face, chest, shoulders, or back. Acne can range from mild to severe and may cause emotional distress and scarring in some cases.
**Causes:**
1. **Excess Oil Production:** Hormonal changes during adolescence or certain times in adulthood can increase sebum (oil) production, leading to clogged pores.
2. **Clogged Pores:** When dead skin cells and oil block hair follicles, bacteria (usually Propionibacterium acnes) can thrive, causing inflammation and acne lesions.
3. **Hormonal Factors:** Fluctuations in hormone levels, such as during puberty, menstrual cycles, pregnancy, or certain medical conditions, can contribute to acne.
4. **Genetics:** A family history of acne can increase the likelihood of developing the condition.
**Types of Acne:**
- **Whiteheads:** Closed plugged pores.
- **Blackheads:** Open plugged pores with a dark surface.
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- **Pustules:** Pimples with pus at their tips.
- **Nodules:** Large, solid, painful lumps beneath the surface.
- **Cysts:** Painful, pus-filled lumps beneath the surface that can cause scarring.
**Treatment:**
Treatment depends on the severity and type of acne but may include:
- **Topical Treatments:** Such as benzoyl peroxide, salicylic acid, or retinoids to reduce bacteria and unclog pores.
- **Oral Medications:** Antibiotics or oral contraceptives for hormonal acne.
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**Prevention and Management:**
- **Cleanse:** Regularly wash skin with a gentle cleanser.
- **Moisturize:** Use non-comedogenic moisturizers to keep skin hydrated without clogging pores.
- **Avoid Irritants:** Such as harsh cosmetics or excessive scrubbing.
- **Sun Protection:** Use sunscreen to prevent exacerbation of acne scars and inflammation.
Acne treatment can take time, and consistency in skincare routines and treatments is crucial. Consulting a dermatologist can help tailor a treatment plan that suits individual needs and reduces the risk of scarring or long-term skin damage.
Cluster Mapping of Medical Tourism in Turkey and Regional Clustering for Heal...
Cluster Mapping of Medical Tourism in Turkey and Regional Clustering for Heal...Istanbul Beykent University (İstanbul Beykent Üniversitesi)
As the world population is aging, Health tourism has become vitally important and will be increased day by day. Because
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services regionally and internationally. There are some aspects of managing and doing marketing activities in order for
medical tourism to be feasible, in a region called as clustering in a region with main stakeholders groups includes Health
providers, Tourism cluster, etc. There are some related and affecting factors to be considered for the feasibility of medical
tourism within this study such as competitiveness, clustering, Entrepreneurship, SMEs. One of the growth phenomenon
is Health tourism in the city of Izmir and Turkey. The model of five competitive forces of Porter and The Diamond model
that is an economical model that shows the four main factors that affect the competitiveness of a nation and its industries
in this study. The short literature of medical tourism and regional clustering have been mentioned.Pharmacology of Drugs for Congestive Heart Failure
Congestive Heart failure is caused by low cardiac output and high sympathetic discharge. Diuretics reduce preload, ACE inhibitors lower afterload, beta blockers reduce sympathetic activity, and digitalis has inotropic effects. Newer medications target vasodilation and myosin activation to improve heart efficiency while lowering energy requirements. Combination therapy, following an assessment of cardiac function and volume status, is the most effective strategy to heart failure care.
PGx Analysis in VarSeq: A User’s Perspective
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The purpose of this webcast is to:
Discuss and demonstrate the approaches with array and NGS genotyping methods for star allele calling to prep for downstream analysis.
Following genotyping, explore alternative tertiary workflow concepts in VarSeq to handle PGx reporting.
Moreover, we will include insights users will need to consider when validating their PGx workflow for all possible star alleles and options you have for automating your PGx analysis for large number of samples. Please join us for a session dedicated to the application of star allele genotyping and subsequent PGx workflows in our VarSeq software.
Storyboard on Skin- Innovative Learning (M-pharm) 2nd sem. (Cosmetics)
Skin is the largest organ of the human body, serving crucial functions that include protection, sensation, regulation, and synthesis. Structurally, it consists of three main layers: the epidermis, dermis, and hypodermis (subcutaneous layer).
1. **Epidermis**: The outermost layer primarily composed of epithelial cells called keratinocytes. It provides a protective barrier against environmental factors, pathogens, and UV radiation.
2. **Dermis**: Located beneath the epidermis, the dermis contains connective tissue, blood vessels, hair follicles, and sweat glands. It plays a vital role in supporting and nourishing the epidermis, regulating body temperature, and housing sensory receptors for touch, pressure, temperature, and pain.
3. **Hypodermis**: Also known as the subcutaneous layer, it consists of fat and connective tissue that anchors the skin to underlying structures like muscles and bones. It provides insulation, cushioning, and energy storage.
Skin performs essential functions such as regulating body temperature through sweat production and blood flow control, synthesizing vitamin D when exposed to sunlight, and serving as a sensory interface with the external environment.
Maintaining skin health is crucial for overall well-being, involving proper hygiene, hydration, protection from sun exposure, and avoiding harmful substances. Skin conditions and diseases range from minor irritations to chronic disorders, emphasizing the importance of regular care and medical attention when needed.
Allopurinol (Anti-gout drug).pptx
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Gene Expression System-viral gene delivery Mpharm(Pharamaceutics)
Gene therapy can be broadly defined as the transfer of genetic material to cure a disease or at least to improve the clinical status of a patient.
One of the basic concepts of gene therapy is to transform viruses into genetic shuttles, which will deliver the gene of interest into the target cells.
Safe methods have been devised to do this, using several viral and non-viral vectors.
In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patient's cells instead of using drugs or surgery.
The biggest hurdle faced by medical research in gene therapy is the availability of effective gene-carrying vectors that meet all of the following criteria:
Protection of transgene or genetic cargo from degradative action of systemic and endonucleases,
Delivery of genetic material to the target site, i.e., either cell cytoplasm or nucleus,
Low potential of triggering unwanted immune responses or genotoxicity,
Economical and feasible availability for patients .
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There are a wide variety of vectors used to deliver DNA or oligo nucleotides into mammalian cells, either in vitro or in vivo.
The most common vector system based on retroviruses, adenoviruses, herpes simplex viruses, adeno associated viruses.
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Patellar Instability: Diagnosis Management
Pictorial and detailed description of patellar instability with sign and symptoms and how to diagnose , what investigations you should go with and how to approach with treatment options . I have presented this slide in my 2nd year junior residency in orthopedics at LLRM medical college Meerut and got good reviews for it
After getting it read you will definitely understand the topic.
TEST BANK For Brunner and Suddarth's Textbook of Medical-Surgical Nursing, 14...
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The development of nanogold-based cancer therapy could revolutionize oncology by providing a more targeted, less invasive treatment option. This project contributes to the growing body of research aimed at harnessing nanotechnology for medical applications, paving the way for future clinical trials and potential commercial applications.
Cancer remains one of the leading causes of death worldwide, prompting the need for innovative treatment methods. Nanotechnology offers promising new approaches, including the use of gold nanoparticles (nanogold) for targeted cancer therapy. Nanogold particles possess unique physical and chemical properties that make them suitable for drug delivery, imaging, and photothermal therapy.
acne vulgaris -Mpharm (2nd semester) Cosmetics and cosmeceuticals
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- Video recording of this lecture in Arabic language: https://youtu.be/ECILGWtgZko
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- Link to NephroTube website: www.NephroTube.com
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Lentivirus protocol
- 2. Lentivirus is classified as retroviridae. The pre-integration complex of the lentivirus nuclear protein has the characteristics of phagocytosis, and the viral genome translocates to the nucleus, so that it can infect and replicate in the non-mitotic cells. This feature makes it an ideal tool to introduce a gene into mammalian or other animal cells.
- 3. Transient transfection method is now widely used for generation of recombinant lentivirus. Envelope plasmid, packaging plasmid and recombinant lentivirus plasmid cotransfect 293T cells to get producing cells.
- 4. The experimental process of recombinant lentivirus protocol production is as follows. 1. Obtain the target genes by PCR. 2. Choose the corresponding lentivirus vector plasmid according to your requests. 3. Construct recombinant lentivirus plasmid. 4. Identify the insert gene sequence, and then extract recombinant lentivirus plasmids. ...