Monsanto: Extinction

One Gene = One Protein? Not Even Close!

HW Bush taught how to genetically modify organisms at Monsanto

Bayer Builds Latin America's Largest Seed Factory in Chile

Double Game

Transport through a cell membrane

Tuning the Body's Genes to Fight against Health Threats

We should not be surprised at Rockefeller's hand in this as John D. Rockerfeller' father made his fortune selling "patent" medicine.

William Avery "Bill" Rockefeller, Sr. (November 13, 1810 – May 11, 1906) was an American confidence man who went by the alias of Dr. William Levingston.

He worked as a traveling "botanic physician" who sold elixirs.

His sons, John Davison Rockefeller (July 8, 1839 – May 23, 1937) and William Avery Rockefeller, Jr. (May 31, 1841 – June 24, 1922), were Standard Oil co-founders.

Several systems for induction of transgene expression in plants exist.

Initially inducible systems are used in tobacco, rice, tomato and corn.

Inducible systems offer the possibility of deregulating gene expression levels at particular stages of plant development and in particular tissues of interest.

The more precise temporal and spatial control, obtained by providing the transgenic plant with the appropriate chemical compound or treatment, permits analysis of the function of those genes required for plant viability.

Specific mutation of a gene can be achieved by a two-step process.

Introduction of loxP sites around a functionally essential genomic part followed by a cell type-specific Cre recombinase-mediated excision of the loxP flanked sequence.

The same strategy can be used for cell type-specific overexpression of a transgene, when a strong overall expressing promoter is separated from the coding region of a gene of interest by loxP flanked 'STOP' sequences.

In both scenarios, a Cre recombinase transgene provides spatial control.

Once Cre expression has been switched on and recombination has occurred, the resultant change in gene expression is, in most cases, irreversible.

Attorney Dr. Reiner Fuellmich

Accueil | 'Non' au New Deal pour la Nature

NIH Documents About Origins of SARS-CoV-2

The Story Behind The Story Of Covid19

One GMO Nearly Took Down the Planet

Why Are We Being Fed By A Poison Expert?

Study Links GMOs to Over 22 Different Diseases

Bayer Herbicide Causes Autism at Trace Levels

"Being an educated person requires - being a full person, requires a certain ability to deal with dissonance.

You know, I'm Jewish, was raised Jewish.

I brought up my kids in the Jewish tradition.

We celebrate Jewish traditions.

At the same time, I'm a scientist.

And data means an awful lot to me.

I clearly believe in evolution and it's really hard to get away from the undirected nature of change.

The science leaves very little room for purpose in evolution." - Eric Lander

1902 Archibald Garrod described the inherited disorder alkaptonuria as an inborn error of metabolism.

He proposed that a gene mutation causes a specific defect in the biochemical pathway for eliminating liquid wastes.

The phenotype of the disease - dark urine - is a reflection of this error.

1910 The term "pleiotropie" is first coined by Ludwig Plate in Festschrift.

He defined pleiotropy as occurring when "several characteristics are dependent upon ... [inheritance]; these characteristics will then always appear together and may thus appear correlated".

1930 Ronald Fisher in Geometric Model implies locus mutations as being capable of affecting essentially all traits.

1941 Inborn error of metabolism is rigorously proven by George Beadle and Edward Tatum using the simple bread mold Neurospora.

Molds exposed to radiation lose the ability to produce essential nutrients, and this slowed, even stopped the growth of the mold.

Growth could be restored providing mutated mold a specific nutrient.

Hypothesis: mutations inactivate enzyme (protein) needed to absorb nutrients.

If a cell stitches exons together in one way, it makes one protein.

Stitching the exons together in another way, it makes a different protein.

Cells making antigens may mistranslate proteins and creat autoantigens.

95% of the genes in the human genome participate in this process.

Alternative pre-mRNA splicing in neurons

Rbfox proteins regulate splicing of a large multiprotein complex

Relationship between Alternative Splicing and Proteomic Complexity

Splicing regulator PTBP1 controls the activity of the transcription factor Pbx1 during neuronal differentiation

posion apple

Engineered food and your health: the nutritional status of GMOs

Identifying Collateral Damage in Genome Editing

Third GMO Arctic Apple Gets USDA Approval

Kosher Certification Bans All GMO Ingredients


The kill-switch for CRISPR

Crispr isn't Enough: Gene Editing 2.0

What are genome editing and CRISPR-Cas9?

Analysis of CRISPR: frequent unintended DNA changes

CRISPR-Cas9 for Genome Engineering

Gene Editing / CRISPR

Molecular genetics is primarily concerned with the inter-relationship between information macromolecules DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) and how molecules are used to synthesize polypeptides.

As the vast majority of gene expression is dedicated to polypeptide synthesis, proteins are the major functional end-points of the DNA template and account for the majority of the dry weight of a cell.

The term protein is derived from the Greek proteios, meaning 'of the first rank' and reflects the important roles of proteins in diverse cellular functions as enzymes, receptors, storage proteins, transport proteins (tRNA), transcription factors, signaling molecules, hormones, etc.

Proteins are composed of polypeptide molecules which may be modified by the addition of various carbohydrate side chains or other chemical groups.

Like DNA and RNA, polypeptide molecules are polymers consisting of a linear sequence of repeating units, in this case amino acids.

The latter consist of a positively charged amino group and a negatively charged carboxylic acid (carboxyl) group connected by a central carbon atom to which is attached an identifying side chain.

Recombinant Proteins, Native Proteins

Reshaping Nature through Gene Drives

Modified RNA has a direct effect on DNA

Scientists Alter Genetic Code

Charged molecules are highly soluble in water.

Both DNA and RNA are negatively charged (polyanions) because of the organophosphate charges present in their component nucleotides.

Depending on their amino acid composition, proteins may carry a net positive charge (basic proteins) or a net negative charge (acidic proteins).

The hydrogen bonding potential of water molecules means that molecules with polar groups (including DNA, RNA and proteins) can form multiple interactions with the water molecules, leading to their solubilization.

Thus, even electrically neutral proteins are often readily soluble if they contain an appreciable number of charged or neutral polar amino acids.

The linear backbone of a DNA molecule and of an RNA molecule consists of alternating sugar residues and organophosphate groups.

Whereas the RNA molecules within a cell normally exist as single molecules, the structure of DNA is a double helix in which two DNA molecules (DNA strands) are held together by weak hydrogen bonds to form a DNA duplex.

DNA can adopt different types of helical structure.

Genetic information is encoded by the linear sequence of bases in the DNA strands (the primary structure).

Intermolecular hydrogen bonding permits RNA-DNA duplexes and double-stranded RNA formation which are important requirements for gene expression.

Hydrogen bonding can occur between bases within a single DNA or RNA molecule.

3 Myths of Precise Genome Editing

mRNA vaccine delivery using lipid nanoparticles

Survival of transgenic plant DNA in the human gastrointestinal tract

CRISPR gene editing in human embryos wreaks chromosomal mayhem

Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA

Recent strategies and advances in the fabrication of nano lipid carriers

The expression of genetic information in all cells is mostly a one-way system: DNA specifies the synthesis of RNA and RNA specifies the synthesis of polypeptides, which subsequently form proteins.

Because of its universality, the DNA > RNA > polypeptide (protein) flow of genetic information has been described as the central dogma of gene expression molecular biology.

The shape and structure of proteins is a crucial aspect of molecular gene expression and links our understanding of gene expression to the biology of the cell.

While primarily concerned with protein molecules that act on DNA and RNA sequences, such as transcription factors and histones, the study of gene expression also focuses on where in the cell expression is modulated.

In fact, the modulation of gene expression can occur in the nucleus, the cytoplasm, or even at the cell membrane due to the impact of proteins on RNA in those cellular subregions.

In his Nobel lecture, given shortly after he joined the Rockefeller Institute for Medical Research, Edward L. Tatum outlined the concepts fundamental to his one-gene, one-enzyme (understood today as one-gene, one-polypeptide) hypothesis:

all biochemical processes in all organisms are under genetic control;

biochemical processes are resolvable into a series of individual reactions;

each reaction is controlled in a primary fashion by a single gene - 1:1 correspondence of gene and biochemical reaction exists;

mutation of a single gene results only in an alteration in the ability of the cell to carry out a single primary chemical reaction.

Geneticist George W. Beadle and the biochemist Edward L. Tatum were awarded the Nobel Prize largely through the auspices of the Rockefeller Foundation, for this hypothesis which turned out to be entirely false !!!

A cell uses the DNA molecule in the nucleus as a template for protein production.

The cell sends a 'messenger RNA' = mRNA into the nucleus to retrieve the encoding.

The mRNA takes the copied "recipe" out of the nucleus to the ribosome, which is where proteins are made.

In eukaryotic cells (the kinds of cells found in plants and animals), however, something very interesting happens before the mRNA leaves the nucleus.

Some parts of the mRNA are cut away, and the remaining parts are then stitched back together.

The parts of the mRNA that are cut away never leave the nucleus, so they are called introns (they stay IN the nucleus).

Introns regulate the amount of the various proteins that are being made.

"For a while, geneticists didn't know the purpose of introns, so in typical evolutionary fashion, many decided that they had no purpose, and introns were lumped into the category of "junk DNA." As we have learned more about genetics, we have learned that the evolution-inspired idea of "junk DNA" is, itself, junk, although some evolutionists still cling to it." - Jay L. Wile

The remaining parts that are stitched together are called exons (they EXit the nucleus).

Each exon represents a "module" of useful information.

If the cell stitches the exons together in one way, it makes one protein.

If it stitches the exons together in another way, it makes a different protein.
As a result, a single gene can actually produce many different proteins.

ALL of molecular biology is based on this fatal error.

Genetically modified organisms and the industrial manipulation of molecular genetics is based on a theory that has turned out to be entirely false.

Unintended toxic proteins is the transgenic standard !

Physical limits to magnetogenetics

Structural characterization of encapsulated bacterial nanocompartments

Widespread distribution of encapsulin nanocompartments reveals functional diversity

mRNA vaccine delivery using lipid nanoparticles

The promise of activating the humoral and the cellular arms of the immune system has driven the development of DNA vaccines over the last decades.

Live-attenuated vaccines are the most potent in activating both cellular and humoral immunity.

However, these vaccines exhibit considerable safety drawbacks.

Attenuated pathogens have the potential to revert to a pathogenic form.

DNA therapeutics have to reach the nucleus, while for mRNA therapeutics, the cytosol is the target.

DNA and mRNA vaccines share many similarities.

The main difference between the two approaches is the target location for the delivery of the oligonucleotides.

mRNA vaccines have generated significant interest to replace traditional vaccines due to a number of important attributes that they possess.

mRNA vaccines elicit a potent immune response including antibodies and cytotoxic T-cells.

Successful cytosolic delivery of mRNA, encoding for an antigen, results in vaccine epitope synthesis of the transfected cells.

As a result, mRNA therapeutics are easier to deliver as they do not have to cross the nuclear membrane.

mRNA synthesis and purification are fast, easy and low cost in comparision, even though mRNA is highly unstable under physiological conditions.

Several strategies have been developed for RNA delivery, including RNA conjugates, modified RNA, viral vectors, microparticles and nanoparticles.

Viral vectors are the obvious choice for delivery, as virus have naturally evolved to become highly efficient at nucleic-acid delivery.

Lipid nanoparticles (LNPs) are among the most frequently used vectors for in vivo RNA delivery.

In addition to ionizable cationic lipids, phospholipids, cholesterol and lipid anchored polyethylene glycol (PEG) are the most commonly used components for LNP formulations.

Lipids are fatty acids insoluble in water but soluble in organic solvents.

Lipids come in four forms natural oils/fats, waxes, phospholipids and steroids.

The theory of vesicle formation assumes that LNP formation is based on disk-like bilayered fragments whose edges are stabilized by ethanol.

Phospholipids play a structural role in LNPs helping with the formation and disruption of the lipid bilayer to facilitate endosomal escape.

Furthermore, some phospholipids possess polymorphic features and promote a transition from a lamellar to a hexagonal phase in the endosome.

The properties of individual LNPs strongly depend on local, microscopic mixing rates; diffusive transport effects can lead to LNPs with variable compositions.

Early synthesis methods relied on the formation of micrometer-sized vesicles by suspending lipids in water, followed by sonication to produce submicrometer sized particles.

This top–down approach has many limitations, including molecular degradation, contamination and lack of scalability.

Extrusion of a lipid film through a small filter has been a popular synthesis method, often used at the laboratory scale using syringe miniextruders.

Other synthesis methods include the condensation of a lipid ethanol solution by rapid injection into a vigorously stirred aqueous buffer.

Newer synthesis methods directly mix the lipid–ethanol phase with an aqueous solution of mRNA in a small T-piece; flow, mixing rates, controlled by pumps.

In this way, LNPs with diameters of 70 nm or larger and high encapsulation efficiencies can be generated.

Decorating the LNPs with immune cell receptors may facilitate the uptake by the desired type of immune cells.

The most important targets for mRNA vaccines are antigen presenting cells (APCs), with dendritic cells (DCs) likely being the most relevant cell type.

APCs are concentrated at high density in lymph nodes (LNs).

The theory is transfected DCs express the mRNA-encoded antigen.

The antigens are subsequently processed by the proteasome, and the generated peptide epitopes enter the endoplasmic reticulum where they are loaded onto major histocompatibility complex (MHC) class I molecules.

The MHC class I molecules are transported to the surface of the cell where the epitopes are presented to CD8 T cells along with costimulatory signals.

Resence of antigen fragments on MHC II induces antigen-specific antibodies.

There is a pathway for the presentation of protein antigens on MHCI, not yet fully understood, often too weak to elicit a potent cytotoxic immune response.

Aluminum salts, used to enhance the immune response of traditional vaccines, thought to be related to the effect of prolonged antigen exposure, is still not understood in detail.

Including adjuvants with the LNPs provides a way to further increase the potency of the vaccine and guide the immune response in the desired direction.

In order to mount a strong adaptive immune response, a vaccine needs to reach the LNs, where T lymphocyte activation and proliferation occurs.

Affinity maturation and isotype switching of antibodies takes place in germinal centers in the LNs.

Intradermal (ID) injection delivers LNPs directly into the skin, an organ which is densely populated with Langerhans cells in the epidermis and with multiple DC subtypes in the dermis.

The ID route of administration has been shown to effectively induce a Th1 type immune response and cytotoxic T lymphocyte induction for mRNA–LNP vaccines.

IV injections of LNP–mRNA vaccines are less common because of the potential of systemic side effects.

Injecting immunogenic material in the blood stream may lead to massive cytokine production, a cytokine storm, that can lead to shock and death.

The thickness of polyethylene glycol (PEG) coating on the LNPs is critical.

Coating the particles with a lipid-anchored PEG containing lipid can reduce complement activation.

PEG coating strongly influences the properties of the LNPs and has to be tailored carefully.

A higher PEG content usually increases the blood circulation time of LNPs, while reducing cellular uptake and interaction with the endosomal membrane.

The surface of an LNP may be decorated with specific targeting sequences which help with homing and subsequent uptake.

Self-amplifying mRNA has been used to prolong protein expression and to increase the immunogenicity of mRNA vaccines, which leads to a dramatic decrease in the effective dose compared with nonreplicating mRNA.

Self-amplifying mRNAs, also termed replicons, are based on retrovirus where the structural viral proteins are replaced with suitable mRNA encoding antigens, as well as with RNA polymerases for RNA replication.

The most studied replicons are derived from alphavirus and flavivirus.

When introduced into the cytosol of cells, the mRNA will express the heterologous genes and replicate.

Through mRNA amplification, large amounts of desired antigens can be synthesized, accounting for up to 20% of total cell protein.

Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines

Infection of cells by coronavirus is effected through the spike glycoprotein.

The coronavirus that causes severe acute respiratory syndrome infects cells expressing the receptor angiotensin-converting enzyme 2.

Here we show that codon optimization of the SARS-CoV spike glycoprotein gene substantially enhanced spike protein expression.

Two retrovirus, simian immunodeficiency virus and murine leukaemia virus, both expressing green fluorescent protein and pseudotyped with SARS-CoV spike glycoprotein variants, infected HEK293T cells stably expressing ACE2.

Infection mediated by an spike glycoprotein variant whose cytoplasmic domain had been truncated and altered to include a fragment of the cytoplasmic tail of the human immunodeficiency virus type 1 envelope glycoprotein was substantially more efficient than that mediated by wild-type spike glycoprotein.

SARS-CoV-2 variants, spike mutations and immune escape

SARS-CoV is not closely related to any of the three previously defined genetic and serological coronavirus groups, although it may be distantly related to group 2 coronavirus.

SARS-CoV spike glycoprotein, a surface glycoprotein facilitating coronavirus entry into receptor-bearing cells, is distinct from other coronavirus.

The gene encoding the spike glycoprotein of SARS-CoV contains many codons used infrequently in mammalian genes for efficiently expressed proteins.

We generated a codon-optimized form of the spike glycoprotein gene and compared its expression with the spike glycoprotein gene of the native viral sequence.

spike glycoprotein was readily detected in HEK293T cells transfected with a plasmid encoding the codon-optimized spike protein.

No spike glycoprotein was detected in cells transfected with a plasmid encoding the native spike glycoprotein gene.

When transfected cells were infected with recombinant vaccinia virus expressing T7 polymerase, which can transcribe message in the cytoplasm, spike glycoprotein was efficiently produced from plasmids containing either codon-optimized or native genes.

The codon-optimized gene expressed more than twice as much spike glycoprotein as the native viral sequence.

Spike glycoprotein can be efficiently expressed from the codon-optimized plasmid without T7 polymerase, we used this plasmid in subsequent studies.

Kary Mullis on Fauci

Vaccine observer says spike glycoprotein is dangerous 'toxin'

spike glycoprotein Efficiently Infect Cells Expressing Angiotensin-Converting Enzyme 2

In the quest for the development of pharmacological switches that control gene expression, no system has been reported that regulates at the translational level but several systems have been constructed:

-Tetracycline-inducible transgenic systems [tetracycline transactivator (tTA) or 'Tet-Off' and reverse tetracycline transactivator (rtTA) or 'Tet-On'] allow for reversible temporal regulation of transgene expression .

Between these, rtTA is better suited for rapid induction of gene expression.

-To permit small-molecule control of transgene translation a farnesyl transferase inhibitor-responsive translation initiation factor was constructed.
This artificial protein is a three-component chimaera consisting of the ribosome recruitment core of the eIF4G1 eukaryotic translation initiation factor, the RNA-binding domain of the R17 bacteriophage coat protein and the plasma membrane localization CAAX motif of farnesylated H-Ras.

This membrane-delocalized translation factor is inactive unless liberated in the cytosol.

Farnesyl transferase inhibitor FTI-277 prevents the membrane association of the CAAX motif and thus increases the cytoplasmic levels of the eIF4G fusion protein, which is then capable of inducing translation of the second cistron of a bicistronic messenger RNA containing an R17-binding site in its intercistronic space.

- The concept of cisgenesis could become a promising approach in future apple breeding.

However, cisgenesis depends on the availability of effective tools for the production of marker-free genetically modified plants.

The development of such plants was recently shown to be possible using a heat shock inducible Flp/FRT recombinase based transformation system allowing the excision of the marker gene from the genome of genetically modified apple plant tissue.

- A new laser mediated method heat shocks groups of cells allowing precise spatio-temporal control of gene expression without requiring knowledge of specific enhancer sequences.

-The baculovirus Autographa californica nucleopolyhedrovirus (AcNPV) has been widely used to achieve a high level of foreign gene expression in insect cells, as well as for efficient gene transduction into mammalian cells without any replication.

In addition to permitting efficient gene delivery, baculovirus has been shown to induce host innate immune responses in various mammalian cells and in mice.

Adventure Thru Inner Space

Adventure Through Inner Space Full Virtual Ride-thru

The major barrier to the clinical application of adenovirus gene therapy for diseases that require stable transgene expression is the immunogenicity of recombinant adenovirus, which ordinarily limits the duration of its effects to a period of about 2 weeks.

If tolerance to adenovirus could be induced then transgene expression could be prolonged if T lymphocytes underwent thymic selection in the presence of adenovirus antigens.

The ability to achieve unresponsiveness to a recombinant adenovirus after inoculation of the thymus in neonates extends the paradigm of intrathymic tolerance induction.

T lymphocytes are exquisitely poised to respond rapidly to pathogens and have proved an instructive model for exploring the regulation of inducible genes.

Individual genes respond to antigenic stimulation in different ways, and it has become clear that the interplay between transcription factors and the chromatin platform of individual genes governs these responses.

Understanding of the complexity of the chromatin platform and the epigenetic mechanisms that contribute to transcriptional control has expanded dramatically in recent years.

These mechanisms include the presence/absence of histone modification marks, which form an epigenetic signature to mark active or inactive genes.

These signatures are dynamically added or removed by epigenetic enzymes, comprising an array of histone-modifying enzymes, including the more recently recognized chromatin-associated signalling kinases.

In addition, chromatin-remodelling complexes physically alter the chromatin structure to regulate chromatin accessibility to transcriptional regulatory factors.

The advent of genome-wide technologies has enabled characterization of the chromatin landscape of T lymphocytes in terms of histone occupancy, histone modification patterns and transcription factor association with specific genomic regulatory regions, generating an image of the T lymphocyte epigenome.

Don't Blame the Bat!

The Remarkable Doctor A. Fauci

Vaccine Induced Disease Enhancement

unique library index

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