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TypeBlog PostPublishedJune 3, 2025, 1:15 p.m....characterized these tools and validated their specificity in mice, non-human primates, and human tissue. Figure...recombinant antibodies against three of the six human glypican proteins (GPC1, GPC3, and GPC4). All IPI...we've already taken them for a test drive! We validated viral packaging and strong expression after inserting...from the antibody pages or in the Addgene Antibody Data Hub. Perhaps these antibodies are just the tools...
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The 12 Days of CRISPR
TypeBlog PostPublishedJan. 3, 2023, 2:15 p.m....activation and interference screens for primary human T cells🎵 On the 3rd day of CRISPR, Addgene gave... On the 5th day of CRISPR, Addgene gave to me: updated CRISPR101 blog posts!🎵 On the 4th day of CRISPR...High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effects🎵 On the 1st ... -
What's New in CRISPR - November 2021
TypeBlog PostPublishedNov. 18, 2021, 2:15 p.m....effective in genome editing in both bacteria and human cells using various delivery methods, including ...RNAs, such as Cre mRNA or Cas9 mRNA and sgRNA, in human and mouse cells. They further develop a version ... AsCas12f1 plasmids Selective endogenous encapsidation for cellular delivery of RNAs By Mike Lacy The... in an approach called Selective Endogenous eNcapsidation for cellular Delivery (SEND). They demonstrate...engineered pegRNAs (epegRNAs) protect the RNA from degradation by cellular exonucleases and broadly improve ... -
Supporting Rare Disease Research with the 2020 BeHEARD Awards
TypeBlog PostPublishedSept. 8, 2020, 2 p.m....collection consists of human and murine solute channels, the team will be able to validate their findings in...in mammalian cells with both human and murine targets thus informing potential for success in a patient-mutation...vectors from Addgene, the lab will express these candidate genes in TS iPSCs to identify whether any rescue... use CRISPR plasmids to generate deletions of candidate genes in euploid iPSCs. “The BeHEARD Addgene reagents... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog PostPublishedJuly 11, 2017, 2:30 p.m....designed by different academic labs, targeting mouse, human, Drosophila melanogaster, and T. gondii are available...contains most of the full genome knockout human and mouse libraries available through Addgene. This means...including other libraries. As of now I have focused on human and mouse SpCas9 knockout libraries, but there are...secondary, CRISPR screen could be used to validate the list of candidate genes. Green Listed, as of now, contains...groups and, as a consequence, which genes are good candidates for being involved in the studied phenotype. ... a massive amount of descriptive transcriptomics data. Custom CRISPR screens designed to target differentially...genes can be the next stage in making sense of such data. These targeted screens can allow a researcher to... -
Viral Vectors 101: Viral Vector Elements
TypeBlog PostPublishedJuly 25, 2023, 1 p.m....infrequent genome integration at the AAVS1 locus in humans. Cap is a structural capsid protein that helps ...-293) during production. Fun fact: There are 57 human adenovirus types, but most vectors are based on ...used to package lab grown viruses. It can be intimidating at first to look at so many plasmid maps and... -
All in a Twist: dsRNA
TypeBlog PostPublishedFeb. 2, 2023, 2:15 p.m....is important for therapeutic delivery, viruses, human disease, and much more! Resources and References...References Wang, I. X., Grunseich, C., Fox, J., et al., Human proteins that interact with RNA/DNA hybrids. Genome...Transcription, translation, and prone to degradation – those are the words that describe RNA! Double ... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 8, 2017, 3:15 p.m....Species of Gene or Insert Choose from: H. sapiens (human), M. musculus (mouse), R. norvegicus (rat), G. gallus...plasmid will produce anything dangerous or toxic to humans in bacteria, select “yes” in the drop-down and ...Table 1: Guidelines for the data fields in the deposit spreadsheet Data Field Description Plasmid...plasmids! We update the spreadsheet every so often and this ensures you are using our most up-to-date version...taken care of and, once we have all of your plasmid data, we’ll send you prepaid shipping materials (i.e....have the most involvement with Step 1, and, while data entry can be a chore, we’ve made it easy to send...plasmid information in a deposit spreadsheet. The data entry process On the Addgene home page, if you scroll... -
Viral Vectors 101: An Introduction to AAV
TypeBlog PostPublishedMay 30, 2023, 1:15 p.m.... 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 PostPublishedJan. 31, 2019, 1:32 p.m....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 PostPublishedAug. 16, 2018, 12:40 p.m....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 PostPublishedJune 1, 2023, 1:15 p.m....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... -
Genome-wide Screening Using CRISPR
TypeBlog PostPublishedAug. 18, 2015, 2:30 p.m....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 PostPublishedNov. 28, 2017, 2 p.m....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 PostPublishedMay 22, 2015, 4:58 p.m....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 PostPublishedMay 18, 2021, 1:15 p.m....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... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog PostPublishedFeb. 7, 2019, 2:16 p.m....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 PostPublishedDec. 15, 2022, 2:15 p.m....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 ... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog PostPublishedSept. 6, 2018, 12:02 p.m....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 PostPublishedFeb. 11, 2025, 2:15 p.m....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...