We narrowed to 302 results for: human dat
-
TypeBlog Post... an organism. These features make AAV ideal for human therapies and in vivo laboratory experiments ranging...medium during adeno-associated virus manufacturing." Human gene therapy 21.10 (2010): 1251-1257. PubMed PMID...we recommend checking the literature and our AAV data hub to see if others have used relevant viruses ... guest blogger Didem Goz Ayturk with edits and updates from Addgenies Karen Guerin and Susanna Stroik....
-
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog Post...secretion of growth promoting proteins, such as human growth factor, vascular endothelial growth factor...early tumor recurrence and poor clinical outcome in human breast cancers." The American journal of pathology...metabolism. Cancer cells promote oxidative stress leading to the degradation of one key enzyme (among others... -
RaPID Detection of RNA-protein Interactions
TypeBlog Post...replication. Human genetic disease. RaPID can also be used to test how RNA variants associated with human disease...then subtracted from Ramanathan et al.’s RaPID datasets. Applications of RaPID RaPID sounds like a nifty...patients. By digging into the literature and analyzing data from The Cancer Genome Atlas, Ramanathan et al. ... -
Viral Vectors 101: AAV Variables That Matter
TypeBlog Post...in a mouse may not work as well in a rat or non-human primate. Likewise, in vitro efficacy doesn’t always...Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089...Expression mediated by three partial sequences of the human tyrosine hydroxylase promoter in vivo. Molecular...virus, a successful AAV experiment starts with validation and optimization. Although there are different...find it is usually faster and easier to do the validation first. AAV-mediated gene expression can differ...inducing drug, so be sure to test for this when validating your viral vector. Titer Once you have selected...measurements. The number itself is determined through validation and optimization of your expression system for... -
TALEN Guide
TypeCollection...investigating gene therapy options for a rare disease in humans, TAL effectors could become a key tool in your ... TAL effector targets. The site, which will be updated soon with more features, is already receiving hundreds... -
Genome-wide Screening Using CRISPR
TypeBlog Post...Addgene carries CRISPR libraries that target mouse, human, fly, E. coli, and T. gondii genes. 2) What genetic... of sub-libraries targeting specific classes of human genes. What are the steps involved in a CRISPR screen...This post was updated on Aug 20, 2020. What genes are important in your phenotype of interest? Many scientists...Each hit from the screen should be independently validated to ensure that the desired modification produces...place. With the proper experimental design and validation practices, CRISPR libraries can help you learn... -
Transferable Skills Guide: Teamwork
TypeBlog Post...the most extraordinary collection of talent, of human knowledge, that has ever been gathered at the White...Thomas Jefferson dined alone.” There was a time in human history when an intelligent, motivated person really...articles in the English version of Wikipedia alone. Humans have been forced to specialize in order to deal...finite goals. For example, when it came time to update how we make and distribute kits, I helped assemble...with the help of these people and others, we have updated our kit process substantially and have plenty of... -
Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards
TypeBlog Post...easy-to-implement confinement strategies that are robust to human error. Used in combination with standard barrier... shown that barrier protocols are vulnerable to human error. And with gene drives, one mistake can be ...first drive. Laboratories interested in building candidate gene drives intended for eventual release should...sake of laboratory convenience. Disclaimer: Recommendations concerning the appropriate degree of confinement... -
Hot Plasmids and Viral Preps - May 2021
TypeBlog Post...antibody toolbox, they screened antibody candidates using a synthetic human scFV library and characterized them...SARS-CoV-2 infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface...slow off-rate binders. The best candidates were validated by IP-MS using HEK293 cell lysate where the ...al., 2021. After validating this method through monitoring GFP degradation, this protein compartment...https://doi.org/10.1126/science.abf9302 Validated recombinant antibody toolbox for aminoacyl tRNA... (aaRS) have been created, characterized, and validated by the Gräslund lab. Recent findings suggest that...inflammation and autoimmune diseases, so these validated antibody tools could be used to study the role... -
Fluorescent Protein Guide: Subcellular Localization
TypeCollection...Targeting Gene/Signal Fluorescent Protein PI 71367 Human Beta-catenin GFP Adherens Junctions Beta-catenin...Autophagosome LC3 EGFP Tamotsu Yoshimori 24920 pEGFP-LC3 (human) Autophagosome LC3 EGFP Toren Finkel 14435 mRFP-...Lck-mTurquoise2 Plasma Membrane Lck N-terminal lipidation signal mTurquoise2 Dorus Gadella 98821 Lck-mScarlet-I...Lck-mScarlet-I Plasma Membrane Lck N-terminal lipidation signal mScarlet-I Dorus Gadella 50057 pLYS1-FLAG-... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...Leonetti to tag all 22,000 protein coding genes in the human genome with fluorescent proteins. For each gene,...mutating. (Note: the most common synonym in the human genome is chosen for a silent mutation.) How to... much as possible, it uses standard tools and databases, including primer3, biopython, bwa, samtools, ...re-cutting when needed. It gives you control over your data. CrispyCrunch keeps a record of all your designs...control. The only ones we found that worked with NGS data were: Crispresso CrispRVariants AmpliCan We chose...Cut-to-insert distance was fitted to experimental data of its effect on HDR efficiency. These values (...reagents Perform your wet lab experiment Analyze the data For more detailed information on using CrispyCrunch... -
8 Great Depositor Research Stories
TypeBlog Post...3,000 antibodies for use in mice and human brain IHC and sharing the data on the mAb3D atlas website. The...Guest blogger Abhi Aggarwal gets the reader up-to-date on the latest version of the iGluSnFR glutamate ... -
Fluorescent Protein Guide: Biosensors
TypeCollection...functionally distinct interneurons in mice, non-human primates and humans. Nat Neurosci. 2020 Dec;23(12):1629-1636...action of redox-directed cancer therapeutics using a human peroxiredoxin-2-based probe. Nat Commun. 2018 Aug...pHluorin-mKate2, Tandem Fluorescent Protein-Tagged Human LC3 for the Monitoring of Mammalian Autophagy. PLoS...Caveolae respond to acute oxidative stress through membrane lipid peroxidation, cytosolic release of CAVIN1...cellular compartments or organelles. Addgene’s database includes many fluorescent biosensors that signal...Cytosolic or mitochondrial sensor for H 2 O 2 oxidation (roGFP2-Orp1) Multiplexed Optical Sensors in Arrayed...targeted DsRed1-E5 as a reporter for mitochondrial oxidative stress A novel MitoTimer reporter gene for mitochondrial... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...Geobacillus stearothermophilus is more stable in human plasma than the standard spCas9 (Harrington et al..."A thermostable Cas9 with increased lifetime in human plasma." Nature Communications8.1 (2017): 1424. ...are quickly cleared from the cell via protein degradation pathways. This makes Cas9 RNPs useful for CRISPR... Addgene.org Browse all CRISPR plasmids Find validated gRNAs Check out our CRISPR guide page ... -
Degrading DNA with Cascade-Cas3
TypeBlog Post...induces broad and unidirectional genome editing in human cells. Nature Communications, 10(1). https://doi.org... I CRISPR–Cas systems for genome engineering in human cells. Nature Biotechnology, 37(12), 1471–1477. ...binding and subsequent DNA degradation appears to be conserved. This DNA degradation portion can be bypassed.... These alterations are achieved through DNA degradation, thanks to the unique capabilities of Cas3 and...of DNA, Cascade caught up to it, and then DNA degradation continued. However, Cas3 cannot bind DNA on its...Hopefully more people will want to utilize Cas3’s degradative power in the future! References and Resources.... (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3. Science, 361(6397). https... -
Viral Production at Addgene
TypeBlog Post...viral vector genome titers by droplet digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://...Infectious Titer by a Novel Droplet Digital PCR Method. Human Gene Therapy Methods, 29(2), 96–103. https://doi.org...at the Addgene blog Find and share AAV Data with Addgene's Data Hub Digital Droplet for AAV Quantification... the absence of unexpected DNA contaminants. The data is also used to determine recombination rates in... vivo for gene expression and/or function. These data are sometimes reported or posted on the material...supernatant is tested for mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination...written by multiple Addgenies and most recently updated by Ina Ersing in August 2023. Resources and references... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog Post...mammalian cells, but mammalian cells can perform human-like post-translational modifications that bacterial...production system, cell lines typically used include Human Embryonic Kidney (HEK) cells and Chinese Hamster...performed during your harvest. Validation Recombinant antibodies should be validated to ensure they bind their...antigens specifically. Possible applications for validation include western blot, immunohistochemistry, and...produced, harvested, purified, quantified, and validated, it’s ready for use! You can learn more about ... -
Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus
TypeBlog Post... along with other viruses that cause disease in humans such as measles, mumps, and rabies. Research on...for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome...VP30, and L (reviewed in 5). The RNA genome is encapsidated by the nucleoprotein, NP. NP associates with...mRNAs are translated) by cellular machinery. NP encapsidates the negative sense, single-stranded RNA minigenome...viral polymerase complex then recognizes the encapsidated minigenome as a template for replication and... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog Post... reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. (2016). PubMed PMID: 27272384...Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells." Nat Biotechnol. (2016). PubMed PMID: 27347757...Updated Aug 13, 2020. Note: Cpf1 is also called Cas12a. In 2015, Zetsche et al. added to the CRISPR ...activity in mammalian cells. From 16 diverse Cpf1 candidates, Zetsche et al. found two that display robust...publication at Addgene. Makarova, Kira S., et al. "An updated evolutionary classification of CRISPR-Cas systems... -
Healthcare Consulting: A Door to the Business of Life Sciences
TypeBlog Post...whether the client invests substantial monetary and human resources towards the development of a new therapy...the business of life sciences and it's impact on human health. Resources on the Addgene Blog Learn... and help resolve it through well-informed recommendations. Healthcare consulting firms that specialize...designing a strategic vision for a portfolio of candidate drugs. A decision like that would allow the client...especially helpful since they both provide up-to-date information about the industry, recruiting processes...