...detectable non-specific activity. Its major advantage is the significantly reduced amount of off-target toxicity...enzyme can lead to off targets and result in significant, negative effects in RNA targeting experiments...using Prime editing alongside some creative (and significant!) protein engineering. They combined Prime editing...The potential of CRISPR has yet to fully taken advantage of, but exciting new tools like these represent...

...thought to activate programmed cell death or a dormant state for bacterial cells that have been infected... Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They characterized...displays similar knockdown efficiency but no significant off-target effects, a distinct contrast to the... In short, the Cas13a molecule brings a much anticipated element of diversity to the CRISPR toolbox and...Check Out Our CRISPR Featured Topic Page Learn about Anti-CRISPRs Multiplex Genome Editing with Cpf1 Resources...

...viral genome and the absence of unexpected DNA contaminants. The data is also used to determine recombination...properties of the final vector preparation, the quantity of gram-negative bacterial endotoxin is ensured...within representative vector preparations was quantified with electron microscopy after negative staining...Approximately two weeks post-thaw, cell culture supernatant is tested for mycoplasma contamination. To date... Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors 101: An Introduction to AAV References...

...were less significant than when non-targeting guides are used as controls. Less significant p-values lead...genome-wide screens.   Figure 1: Summary of the advantages of the new approach to genome-wide CRISPR screen... Screens Before diving into the results, it’s important to note that in Morgens et al., guides were considered...of on-target activity. However, there was no significant difference in depletion of ricin sensitizer genes...

... comments on the AMA process below, but we also wanted to share some thoughts on why we decided to do ...Software for Biology) on Reddit and got some substantial traffic that we thought it would be a good idea...worried about selling yourself and really do just want to get information about yourself and your work ...internet content for scientists in training is abundant, helpful, and sometimes excellent. Finally, as...as I said in the AMA, it’s incredibly important to find good mentors and incentivize good mentorship: ...

...volume of lower titer virus. In such cases you may want to consider concentrating your viral prep. Storage...al. found that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, ...protein attaches to cell surface receptors, other important binding events take place during the course of...source. Labs that routinely produce lentivirus may want to consider testing a variety of cation and serum...Ward T.H., Margison G.P. “Mycoplasma infection significantly alters microarray gene expression profiles.”...

...thanks to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is a PhD candidate...This post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-...cut will continue to flow and be captured by an antibody further along on the strip. Highlights: Of the... cDNA. Isothermal amplification is done at one constant temperature, eliminating the need for thermocyclers...

...should be supplied (Baloban et al., 2017).  It is important to consider effective brightness of NIR FPs in...molecular brightness (a product of the fluorescence quantum yield and the extinction coefficient) (Shcherbakova...They were engineered to incorporate BV. Their advantages are their small size (17 kDa for miRFPnanos, ... highlighted in blue. aExtinction coefficient. bQuantum yield. cDetermined in mammalian cells. dDetermined... near-infrared proteins provide a scaffold for antigen-dependent fluorescent nanobodies. Nat Methods submitted...

...but it’s also important not to load yourself up with too many tasks. If you’re constantly switching between... recommend the book, “Bored and Brilliant” for learning more fantastic reasons to remove yourself from...ll discuss in the planning section below, it’s important to dedicate specific amounts of time to specific...

...good article at NatureJobs: Interviews: The All-Important Job Talk. 1. Watch other people give great talks...Think Keystone Keynote presentation or Ted Talk. You want to give the audience something to think about, argue... time to prepare (and it does take time). For important talks, ask your peer mentors to check your slides...Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than too long, and...to practice and over time you can master this important science skill. Additional Resources Joanne'...

...CRISPR tools for use in plants.  Listen to the segment on CRISPR tools for plants. Special thanks to our...enhancing iPSC production, and CRISPR tools for plants. You can find additional hot plasmids in our quarterly...: Interview with Darrell Kotton Find additional plant biology blog posts Resources at Addgene.org Sign...

...CRISPR, Addgene gave to me: Functional Genetic Variants Revealed by Massively Parallel Precise Genome ...prime editing systems by manipulating cellular determinants of editing outcomes🎵 On the 5th day of CRISPR...CRISPR, Addgene gave to me: High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral...prime editor with enhanced editing efficiency in plants 🎵 ...

...photo wearing holiday sweaters and Santa hats. There was even a giant taco... Miller lab annual #DeckTheLab...their door. What a fun video of Mario, dressed as Santa, delivering gifts to the Mushroom Kingdom. Kravitz.... Getting in the spirit for the holiday season. Santa frog is coming to town. @Addgene @UMich_MCDB #christmastaco...

...expression and largely defines the amount of recombinant protein that will be produced. Many commonly-...Tet-Off vectors (Tet-Advanced)” are the most recent variants of the Tet-Off system.  For more information on...the first 24 hours of culture, when glucose is abundant and ethanol concentrations are low, pADH1 displays...pADH1 activity. Researchers have engineered pADH1 variants that are less sensitive to ethanol, including ... transcription from the UAS promoter. This is important because it allows them to be used together for...

...enable sorting and selection of transduced cells. Grant Challen’s lab also created constitutive (pCMV-dCas9...fused LSD1 to x-Cas9(3.7). x-Cas9(3.7) is a SpCas9 mutant that exhibits increased PAM flexibility, and this...recruit epigenetic machinery - but is there an advantage to using direct epigenetic modifiers? As with ...will determine the tool that you should use. If you want to study the effects of one particular modification...tool would be your best bet. Another potential advantage of CRISPR epigenetic tools is their persistence...

...Goldilocks Antibody: Guest blogger Ava Shipman explains how the Wu lab is screening over 3,000 antibodies for... to RUBY-red, a bright red pigment reporter for plants.  Cas7-11 and PASTE: Omar Abudayyeh and Jonathan...your own work!) If you've got a research story you want to share on our blog, please reach out to [email protected]...

...introduction of the model antigen, Ovalbumin, into cell lines to study antigen-specific responses. Find...screens have been used to interrogate T cells, an important immune cell type, in more complex and physiological...for a screen that assessed genes that regulate antigen-specific CD8+ T cell responses to a murine tumor...

...can save you significant time and money. For instance, this report shows that anterograde, monosynaptic...see from the community is your negative data. We want to make sure people are selecting AAVs that will...work was cited by others?)  Ready to submit?  Fantastic! The next 25 submissions will receive a 20% off...

...Instrumental Access participants. Like Seeding Labs, Addgene also feels it’s incredibly important for scientific...challenges. However, these same researchers face significant barriers to scientific discovery and career development...accelerate biology research in the labs they’re granted to, as well as shine a light on some of the great...

...identify what your plasmid contains and other important information. Tip: A lowercase "p" is often used...simple piece of information can often convey many important details. Once you know the backbone a plasmid ... on, you can usually derive: a) the bacterial antibiotic resistance, b) the promoter that drives the insert...position 295 was mutated to an Alanine, Q295A.  Mutant version: pBACKBONE-Flag-hGene(Q295A)-EGFP  Wildtype...