Imponerende arbeid @polygon 
12 Likes
Må bare slutte meg til @sjog.
Om du har rett eller ikke; dette er jobbing og graving på høyt nivå. Gålr dine antakelser fullt inn, er det bare å spenne sikkerhetsbeltene hardt til de nærmeste ukene.
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Krysser fingrene for at ditt sherlock arbeid stemmer.
8 Likes
Långt inlägg…
Det som diskutera flitigast gällande fimaNac är framför allt mRNA baserade mediciner.
Något som inte tas upp lika mycket är CRISPR, gensaxen som många tror kommer revolusionera medicin. Svårigheten ligger som vanligt i leverering 
Från Thermi Fishers hemsida, de producerar, Lipofectamine som används i försök inom in vivo försök med CRISPR.
Transfection is the process by which CRISPR-Cas9 DNA, mRNA and protein systems are introduced into eukaryotic cells. Techniques vary widely and include lipid nanoparticle–mediated transfection, viral delivery, and physical methods such as electroporation. Our Invitrogen Lipofectamine family of reagents paired with the Neon Transfection System offer complete delivery solutions to address your genome editing needs. We have optimized protocols for each product option to help you achieve high cleavage efficiency and the ease of delivery you expect.
Om man sedan går till forskningsartiklar som finns låter det inte lika övertygande om lipofectamine.
Delivery Aspects of CRISPR/Cas for in Vivo Genome Editing. Danny Wilbie et al. Acc Chem Res. 2019.
In this Account, we focus on the delivery aspects of CRISPR/Cas for therapeutic applications in vivo. Safe and effective delivery of the CRISPR/Cas components into the nucleus of affected cells is essential for therapeutic gene editing. These components can be delivered in several formats, such as pDNA, viral vectors, or ribonuclear complexes. In the ideal case, the delivery system should address the current limitations of CRISPR gene editing, which are (1) lack of targeting specific tissues or cells, (2) the inability to enter cells, (3) activation of the immune system, and (4) off-target events.
fimaNac gör det möjligt att 1: aktivera lokalt, 2: ökar möjligheten att komma in i cellerna/cytoplasman och 3: därmed få önskad effekt där man önskar och samtidigt minska oönskade effekter på andra platser.
To circumvent most of these problems, initial therapeutic applications of CRISPR/Cas were performed on cells ex vivo via classical methods (e.g., microinjection or electroporation) and novel methods (e.g., TRIAMF and iTOP). Ideal candidates for such methods are, for example, hematopoietic cells, but not all tissue types are suited for ex vivo manipulation. For direct in vivo application, however, delivery systems are needed that can target the CRISPR/Cas components to specific tissues or cells in the human body, without causing immune activation or causing high frequencies of off-target effects . Viral systems have been used as a first resort to transduce cells in vivo. These systems suffer from problems related to packaging constraints, immunogenicity, and longevity of Cas expression, which favors off-target events. Viral vectors are as such not the best choice for direct in vivo delivery of CRISPR/Cas. Synthetic vectors can deliver nucleic acids as well, without the innate disadvantages of viral vectors. They can be classed into lipid, polymeric, and inorganic particles, all of which have been reported in the literature. The advantage of synthetic systems is that they can deliver the CRISPR/Cas system also as a preformed ribonucleoprotein complex. The transient nature of this approach favors low frequencies of off-target events and minimizes the window of immune activation.
Moreover, from a pharmaceutical perspective, synthetic delivery systems are much easier to scale up for clinical use compared to viral vectors and can be chemically functionalized with ligands to obtain target cell specificity. The first preclinical results with lipid nanoparticles delivering CRISPR/Cas either as mRNA or ribonucleoproteins are very promising. The goal is translating these CRISPR/Cas therapeutics to a clinical setting as well. Taken together, these current trends seem to favor the use of sgRNA/Cas ribonucleoprotein complexes delivered in vivo by synthetic particles.
Troligtvis kan fimaNac finna en plats även här.
Delivering CRISPR: A Review of the Challenges and Approaches
Christopher A Lino et al. Drug Deliv. 2018 Nov.
Abstract
Gene therapy has long held promise to correct a variety of human diseases and defects. Discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR), the mechanism of the CRISPR-based prokaryotic adaptive immune system (CRISPR-associated system, Cas), and its repurposing into a potent gene editing tool has revolutionized the field of molecular biology and generated excitement for new and improved gene therapies. Additionally, the simplicity and flexibility of the CRISPR/Cas9 site-specific nuclease system has led to its widespread use in many biological research areas including development of model cell lines, discovering mechanisms of disease, identifying disease targets, development of transgene animals and plants, and transcriptional modulation. In this review, we present the brief history and basic mechanisms of the CRISPR/Cas9 system and its predecessors (ZFNs and TALENs), lessons learned from past human gene therapy efforts, and recent modifications of CRISPR/Cas9 to provide functions beyond gene editing.
We introduce several factors that influence CRISPR/Cas9 efficacy which must be addressed before effective in vivo human gene therapy can be realized.
The focus then turns to the most difficult barrier to potential in vivo use of CRISPR/Cas9, delivery.
We detail the various cargos and delivery vehicles reported for CRISPR/Cas9, including physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles), and discuss their relative merits. We also examine several technologies that, while not currently reported for CRISPR/Cas9 delivery, appear to have promise in this field. The therapeutic potential of CRISPR/Cas9 is vast and will only increase as the technology and its delivery improves
.
Flera dödsfall i tidiga försök med genterapi:
It is therefore critical that gene therapy technologies allow for highly specific editing of the genome to reduce the risk of undesired mutagenesis, and that the delivery vehicle allows for safe and efficient transport to the target.
.
fimaNac kan troligen hjälpa till att få önskad effekt med ökad precision, dvs ändra genomet lokalt och inte på oönskade platser i kroppen.
Careful consideration and development of both the gene editing tool and the delivery mechanism will be required if the full potential of therapeutic gene editing is to be realized.
Crispr och lipid nanoparticles.
There are substantial drawbacks for delivery of CRISPR/Cas9 components via lipid nanoparticle. First, there are both external and internal barriers that must be considered. Once the nanoparticle has passed through the surface of the cell, it is typically encased within an endosome. Encased contents can very rapidly be directed by the cell into the lysosomal pathway, causing the degradation of all lysosome contents. Therefore, the cargo must escape the endosome. Also, if the Cas9:sgRNA complex can escape the endosome, it must also translocate to the nucleus, which can also be a potential point of failure.
Because of this, it is rare to see particularly high efficacies when delivery CRISPR/Cas9 components via lipid nanoparticles. While Wang et al. (2016) could achieve ∼70% in vitro modification efficiency in cells (see Figure 7(A)), that only came after an intense screen to determine the most optimal lipids with which to construct their liposomes for their system. Finally, lipid nanoparticles are like virus particles in that the nature and size of the cargo, along with the target cell type, highly affect transfection efficiency and the types of lipids that are appropriate or useful in the system.
Because of their lack of viral components, there will always be interest in improving lipid nanoparticles to deliver CRISPR/Cas9 components. This improvement process can come through the screening of better lipid carriers, as above; better decorations on the liposome surface to help target particles to specific cells or tissues, avoid immune system detection, and facilitate endosomal escape; and improved packaging of CRISPR/Cas9 components, increasing the odds of some subset of packaged molecules to be appropriately delivered.
Other common polymeric vectors for DNA delivery are polyethenimine (PEI) and poly(l-lysine) (PLL). Branched PEI have high charge density, facilitating efficient plasmid DNA packing, and pH-buffering ability which enables escape from endosomes. However, branched PEI is cytotoxic. Therefore, a balance between the desirable properties of branched PEI and the less toxic linear PEI must be struck for effective transfection.
Återigen svår balans mellan effekt och toxicitet.
Refererar till PCIB:s presentation om fimaNac: Enhancement by fimaNac is best under conditions favourable for vehicle safety.
Low concentration of vehicle/nucleic acid complex.
Låg koncentration av den delen som har toxiska egenskaper ger bäst effekt, och det är då den fungerar som bäst. Vanligtvis måste man ligga på gränsen till toxiska reaktioner för att få önskad effekt, här är det tvärt om. Helt otroligt.
Citat från samma presentation som ovan:
fimaNac is especially advantageous in vivo :
- difficult to achieve a high concentration of vehicle/nucleic acid complex in target cells.
- Toxicity may limit the amount of vehicle used.
Det återkommer hela tiden inom genterapi oavsett typ: toxiciteten begränsar. Svårigheten är att leverera!
Mycket tyder på att det nu är löst för i alla fall vissa indikationer.
Över till AZ och CRISPR
Thursday, 29 January 2015
AstraZeneca today announced four research collaborations aimed at harnessing the power of CRISPR, a pioneering genome-editing technique, across its entire discovery platform in the company’s key therapeutic areas. The technology will allow AstraZeneca to identify and validate new drug targets in preclinical models that closely resemble human disease. AstraZeneca will share cell lines and compounds with its partners and work with them to publish findings of its application of CRISPR technology in peer-reviewed journals, contributing to broader scientific progress in the field. The collaborations complement AstraZeneca’s in-house CRISPR programme and will build on the company’s ‘open innovation’ approach to research and development.
CRISPR (clustered regularly interspaced short palindromic repeats) is a genome-editing tool, which allows scientists to make changes in specific genes far faster and in a much more precise way than ever before. The technology has two components - a homing device to a specific section of DNA (guide-RNA) and enzymatic ‘scissors’ that cut DNA (Cas9 nuclease). In the cell nucleus, the guide-RNA sequence directs the Cas9 nuclease to cause double-stranded breaks in the target DNA sequence. By harnessing the cell’s own DNA-repair apparatus, the gene being targeted can be altered either by deleting it, adding nucleotides to it or by turning its activity on or off. In contrast to previous genome-editing techniques, such as zinc-finger nucleases and TALENs, CRISPR is easier to handle in the laboratory.
Dr. Mene Pangalos, Executive Vice President, Innovative Medicines & Early Development, AstraZeneca, said: “CRISPR is a simple yet powerful tool that enables us to manipulate genes of potential importance in disease pathways and examine the impact of these modifications in a highly precise way. By combining the great science from our labs with these world-renowned academic and industry partners, we will be able to integrate this ground-breaking technology into our research and help accelerate the discovery of novel treatments for patients.”
Jag hoppas att vi inte säljer Nac till AZ utan licenserar ut det till mycket högt pris för att sedan kunna licensiera till flera andra BP om resultaten är så övertygande som det verkar. Skulle AZ köpa lösningen på leverans inom genterapi hoppas jag att vi inte säljer Nac för mindre än 100 miljarder. Låter kanske galet men potentiella intjäningen är hissnande om levereringsgåtan nu är löst.
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Takk for langt, spennende og informativt innlegg @Temlor!
Dette som er uthevet her er så til de grader godt å lese, igjen og igjen!
Her har vi en fundamental utfordring for CRISPR/Cas9-teknologien oppsummert. Sammenholder vi dette med det PCIB har full kontroll på, og som PW gjennomgår stort sett på hver presentasjon, nemlig endosomal escape generelt;
og FimaNAc spesielt;
og
så ser vi teknologier som tilsynelatende passer til hverandre som hånd i hanske.
Legg til uheldige bieffekter ved bruk av andre leveringsmetoder (physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles)), og at dette er et av de virkelig store forskningsområdene for tiden, med vanvittig markedsstørrelse, økende fremover, i tillegg til samarbeidet med AZ - key player på området - der samarbeidet skal finne sitt neste steg i løpet av de neste 5 ukene, så ser vi at PCIB, og vi som er aksjonærer, har en utrolig spennende tid foran oss.
Da jeg gikk inn i PCIB var det helt klart FimaChem som var årsaken, med sine sterke og lovende resultater innen Gallegangskreft. Alt annet var icing on the cake. Innen 5 uker kan icingen nå vise seg å bli lagt på før kaken er ferdig. Det blir 5 utrolig spennende uker fremover, og vi sitter på fremste rad!
The future is imminent!
Edit: La inn denne lenken: http://pcibiotech.no/wp-content/uploads/2014/01/Kan-man-levere-medisiner-med-lys.pdf
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Må innrømme at de siste store innleggene påhører meg “semi erotiske” følelser…
Godt jobbet 
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http://pcibiotech.no/wp-content/uploads/2017/05/PCI-Biotech-Q1-2017-Presentation.pdf
se side 19 der CRISPR er med.
PW ble spurt på en kvartalsgjennomgang om PCIB og CRISPR kunne potensielt ha synergier hvorpå PW bekreftet dette.
Ved påfølgende kvartalsgjennomganger så ble CRISPR tatt med på skissen over fimaNAc.
Mener å huske at PW var veldig positiv til at PCIB kunne tilføre CRISPR-teknologien positivt bidrag.
5 Likes
I den meldingen over fra AZ(børsmelding?) står det at de har inngått 4 samarbeid, vil dette da være potensielt 3 konkurrenter til pcib innenfor NAc delen? Vet vi noe om disse?
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Det er forskjell på utvikling av CRISPR og effektivt delivery av CRISPR stupet.
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Samarbetena handlar mer om att lokalisera gener och olika sjukdomar det handlar om stor mängd data. Andra samarbeten handlar om att ta fram cellinjer för prekliniska försök med mera .
Det enda som jag ser kan direkt handla om delivery är samarbetet med Thermo Fisher som producerar lipofectamine som jag pratade om in inlägget ovan.
Vad PW tycker om lipofectamine vs fimaNac vet vi ju redan
De olika samarbetena listas under, direkt från AZ:s hemsida.
AstraZeneca’s research collaborations are with the following institutions:
The Wellcome Trust Sanger Institute, Cambridge, UK
Under the terms of the collaboration with the Wellcome Trust Sanger Institute, research will focus on deleting specific genes relevant to cancer, cardiovascular, metabolic, respiratory, autoimmune & inflammatory diseases and regenerative medicine to understand their precise role in these conditions. AstraZeneca will provide cell lines that can be targeted using the Sanger Institute’s collection of genome-wide CRISPR guide-RNA libraries to generate populations of cells in which defined genes are switched off. Genes will subsequently be identified by next-generation sequencing and cell populations tested to validate the effects of a given gene on a wide range of physical and biological traits.
“The Sanger Institute’s guide-RNA library enables researchers to target genes with incredible specificity,” said Dr. Kosuke Yusa, Member of Faculty at the Sanger Institute. “CRISPR has transformed the way we study the behaviour of cells and now the application of this powerful technology to the search for effective drugs has the potential to benefit patients.”
The Innovative Genomics Initiative, California
The Innovative Genomics Initiative (IGI) is a joint venture between the University of California, Berkeley and University of California, San Francisco. The research collaboration will focus on either inhibiting (CRISPRi) or activating (CRISPRa) genes to understand their role in disease pathology. The IGI and AstraZeneca will work closely together to identify and validate gene targets relevant to cancer, cardiovascular, metabolic, respiratory, autoimmune and inflammatory diseases and regenerative medicine to understand their precise role in these conditions.
“We are excited to pair the IGI’s premier expertise in CRISPR gene editing and regulation with AstraZeneca’s deep experience in therapeutics,” said Jacob Corn, Scientific Director of the Innovative Genomics Initiative. “I’m confident that, working side-by-side with scientists at AstraZeneca, our collaboration will positively impact drug discovery and development to hasten treatments to patients.”
Thermo Fisher Scientific, Waltham, Massachusetts
Under the terms of the collaboration with Thermo Fisher Scientific, a world-leading reagent and instrument provider, AstraZeneca will receive RNA-guide libraries that target individual known human genes and gene families. AstraZeneca can screen these guides against cell lines to identify new disease targets.
“Through this research collaboration with AstraZeneca, Thermo Fisher is helping to accelerate access to cutting edge genome-editing applications for next generation drug discovery. Enabling more relevant disease models will improve target identification and translation to therapeutics,” said Dr. Jon Chesnut, Director of Synthetic Biology R&D at Thermo Fisher Scientific.
Broad Institute/Whitehead Institute, Cambridge, Massachusetts
The collaboration with the Broad Institute and Whitehead Institute will evaluate a genome-wide CRISPR library against a panel of cancer cell lines with a view to identifying new targets for cancer drug discovery.
In addition to the new collaborations, AstraZeneca’s in-house programme is currently adapting CRISPR technology to streamline and accelerate the production of cell lines and translational models that mimic complex genomic and disease-relevant scenarios.
“Application of the CRISPR technology for precise genome editing in recombinant cell lines and in relevant disease models should enable us to identify novel targets, build better test systems for drug discovery and enhance the translatability of our efficacy and safety models,” said Dr. Lorenz Mayr, Vice President, Reagents & Assay Development, AstraZeneca.
The short video above explaining how CRISPR technology works is available in English, Mandarin and Spanish, and a still image illustrating the technology is available.
Download broadcast video and high-resolution image
About The Wellcome Trust Sanger Institute
The Wellcome Trust Sanger Institute is one of the world’s leading genome centres. Through its ability to conduct research at scale, it is able to engage in bold and long-term exploratory projects that are designed to influence and empower medical science globally. Institute research findings, generated through its own research programmes and through its leading role in international consortia, are being used to develop new diagnostics and treatments for human disease. http://www.sanger.ac.uk/
About Innovative Genomics Initiative
The Innovative Genomics Initiative (IGI) was established in early 2014 at the Li Ka Shing Center for Genomic Engineering at the University of California, Berkeley, and is a joint UC Berkeley/UC San Francisco initiative catalyzing and guiding the global effort in both the academic and commercial research communities to unleash the transformative potential of CRISPR/Cas9 technology for positive human impact.
About the Broad Institute of Harvard and MIT
The Eli and Edythe L. Broad Institute of Harvard and MIT was launched in 2004 to empower this generation of creative scientists to transform medicine. The Broad Institute seeks to describe all the molecular components of life and their connections; discover the molecular basis of major human diseases; develop effective new approaches to diagnostics and therapeutics; and disseminate discoveries, tools, methods and data openly to the entire scientific community.
Founded by MIT, Harvard and its affiliated hospitals, and the visionary Los Angeles philanthropists Eli and Edythe L. Broad, the Broad Institute includes faculty, professional staff and students from throughout the MIT and Harvard biomedical research communities and beyond, with collaborations spanning over a hundred private and public institutions in more than 40 countries worldwide. For further information about the Broad Institute, go to http://www.broadinstitute.org.
About Whitehead Institute
Whitehead Institute is a world-renowned non-profit research institution dedicated to improving human health through basic biomedical research. Wholly independent in its governance, finances, and research programs, Whitehead shares a close affiliation with Massachusetts Institute of Technology through its faculty, who hold joint MIT appointments. http://wi.mit.edu
About Thermo Fisher Scientific
Thermo Fisher Scientific Inc. is the world leader in serving science, with revenues of $17 billion and 50,000 employees in 50 countries. Our mission is to enable our customers to make the world healthier, cleaner and safer. We help our customers accelerate life sciences research, solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. Through our four premier brands – Thermo Scientific, Life Technologies, Fisher Scientific and Unity Lab Services – we offer an unmatched combination of innovative technologies, purchasing convenience and comprehensive support. For more information, please visit www.thermofisher.com.
Annan länk från 2018:
AZ and CRUK team up to go all-in on CRISPR
December 10, 2018
AstraZeneca and Cancer Research UK are collaborating to launch a centre of excellence in genetic screening, cancer modelling and big data processing that will develop CRISPR technology to better understand the biology of cancer.
The Functional Genomics Centre will help create biological models that may be more reflective of human disease, as well as advancing computational approaches to better analyse big datasets, with the aim of accelerating the discovery of new oncology medicines.
CRISPR is a gene editing technology that can be used to edit the desired parts of the genome, which makes it possible to identify a DNA sequence, remove it, replace it or add it more easily than ever before to result in the deletion or introduction of specific nucleotide changes into any gene.
Though it is still an emerging field, the first therapies utilising CRISPR are already being reviewed by the FDA, including Vertex’s CTX001.
“CRISPR in its broadest intent has become embedded in the majority of our therapeutic programmes in terms of our ability to build better disease models, help us identify better medicines and now increasingly in functional genomics to help us identify the targets that we’re working on in the first instance,” Steve Rees, AZ’s VP discovery biology, IMED Biotech Unit, told pharmaphorum at a media briefing. “The collaboration with CRUK in the Centre for Functional Genomics will allow us to run 150 of these screens each year from 2020 – half of those will be for AZ and half for CRUK – and this will apply to the majority of our oncology projects to help us identify targets in the first instance but also help us identify the resistance mechanisms to our drugs such that we can make improved medicines that address those resistances.
“In a couple of years’ time I expect that every drug candidate we select to go into the clinic somewhere along the way will have been touched by CRISPR, in terms of optimising the model system that it’s tested in.”
Functional genomics aims to understand the complex relationship between genetic changes happening within DNA and how these translate to cellular changes in disease. Knowing the functional genomic drivers of disease enables scientists to more accurately select the right drug targets and increases the probability of success in the clinic, and Mene Pangalos, AZ’s executive vice president, innovative medicines & early development, said that the collaboration is part of the company’s wider strategy to increase the success rate of its programmes.
“None of our programmes go in the clinic now without being able to demonstrate proof of mechanism – that you’re actually engaging the target, engaging the pathway and engaging the science, so you know that if it fails you’ve tested your hypothesis and you’ve learnt something as a consequence.”
Recently the company had two high-profile late-stage failures with its cancer immunotherapy Imfinzi (durvalumab), in phase 3 trials for head and neck cancer and lung cancer.
At the Centre, scientists will have access to the next generation of CRISPR libraries for silencing or activating every gene in the genome, accessed through an extension of the existing collaboration between AstraZeneca and the Wellcome Sanger Institute.
The centre will be housed in the Milner Therapeutics Institute at the University of Cambridge and operationalised through Cancer Research UK’s Therapeutic Discovery Laboratories – the charity’s in-house drug discovery unit focused on establishing drug discovery alliances with industry.
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PCI Biotech: Disclosure of voting rights for Chairman of the Board
1 Like
PCI Biotech: Disclosure of voting rights for Chairman of the Board
4 Likes
PCI Biotech: Minutes from Annual General Meeting 2020
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Da skal det vel fordeles et antall opsjoner en av de første dagene.
4 Likes
For en som ikke har peiling på slike ting. Kan du gi en kort forklaring?
1 Like
Dette er interessant!
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Nja… Kunne gjerne kommet med noe mer håndfast og gitt eksempler på dett… Blir litt bare en påstand slik det står nå…
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Enig!
Hvilke andre avtaler tenker du på Lenny?
2 Likes
Hver måned kårer jeg det jeg kaller Teksperter™ for noen av de mest populære investeringene våre
Det er de 3 medlemmene som har fått flest likes på innleggene sine de siste 90 dagene. Teksperter™ får også en unikt merke på profilen sin og et trofé-ikon ved siden av navnet sitt. Du kan bli Tekspert™ i flere aksjer/investeringer, og troféet vil bare vises i tråder der du er Tekspert™.
Her er denne månedens Teksperter™ og det mest likte innlegget deres fra de siste 90 dagene:
- @anon50125742 (2603 likes)
- @Jubel (1539 likes)
- @Investor (1388 likes)
Resten av topp 10:
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@polygon (1344 likes)
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@abacus (1307 likes)
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@Savepig (1298 likes)
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@Bonusfrelst (1234 likes)
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@sjog (1221 likes)
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@Longtrader (1194 likes)
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@bamse (1176 likes)
Gratulerer!
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Litt og topic men jeg håper ikke å se meg selv på denne lista neste mnd. Håper ikke de med reell kunnskap og kompetanse slutter å skrive på TI. Selv er jeg kun en lekmann som søker kunnskap her, kommer med egne tanker rundt ting og forsøker meg med noen innspill innimellom.
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