...governments exist in reality and science is how we learn about reality. Formally, a government can be defined...public discourse, many scientists have chosen to leave the lab and march. Unintended consequences While...

...multiplexing gRNAs here). Cas9 is commonly tagged with a nuclear localization sequence to enhance targeting to the...plant species or cell type (Belhaj et al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9...

...loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient Cas9 can bind loci in the genome that...interfere with the function of the protein. This is clearly the case when proteins undergo post-translational...

... 1). In general, the monomeric state of a probe leads to less interference with the oligomeric state of...indicators NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, performance, and...

... in the classroom on our blog. Now that you’ve learned more about popular model organisms in the lab, ... about plasmids in our Plasmids 101 blog series Learn more about CRISPR and genome editing in our CRISPR...

...to specific subcellular locations, such as the nuclear envelope, allowing scientists to probe interactions...expression levels but for many fusion proteins this would lead to protein/organelle aggregation. The Ting lab hypothesized...

...tip: When designing your PCR, ensure there is at least ~200 base pairs of sequence flanking the editing...find a null clone, you should screen, at the very least, 4 clones. Did you use a dual guide system to generate...

...(7). This modification allowed the deletion of release factor 1, the protein responsible for terminating...reference and protocols for use of this system, please reference ‘Rinehart Lab reagents for improved expression...

...fungus Neurospora crassa. The QUAS promoter is less leaky than UAS in the basal state, and co-expression of...expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS, as seen with...

...Mentoring Program Director of Strategic Initiatives. Learn more at https://www.linkedin.com/in/lopezdiegothinkbrain...lopezdiegothinkbrain        Other Resources Visit the IMP website Learn about the Mentor Circle Program at Brigham and Women's...

...time investment. Once the target mRNA is cleaved, the cleaved mRNA can induce further rounds of RNAi. ...

...References 1. Hochedlinger, K. and R. Jaenisch, Nuclear reprogramming and pluripotency. Nature, 2006. 441... Factors that Differentiate the Stem Cell Field Learn how lentiviral vectors can be used for gene delivery...

...used in this approach should also allow for the clear identification of the cell types of interest. By...Kolahdouzan, M., Chen, C. X., You, Z., Benaliouad, F., Gileadi, O., McBride, H. M., Durcan, T. M., Edwards, A....

...like zinc finger nucleases (ZFNs) or transcription-activator-like effector nucleases (TALENs) required...target cleavage, while mismatches toward the 5′ end distal to the PAM often permit target cleavage. Generating...binding. In this formation, its nuclease domains, RuvC and HNH, cleave opposite strands of the target ...popular Cas endonuclease, with many modified versions engineered to increase specificity and nuclease activity...Cas9 The two Cas9 nuclease domains, RuvC and HNH, can be altered to reduce cleavage events to a single...contains mutations in both nuclease domains, D10A and H840A, resulting in no cleavage of target DNA. Cas9n ...increase nuclease fidelity SpCas9-NG - NG; increase in vitro activity SpG - NGN; increase nuclease activity...

...molecular biology enzymes: 5' exonuclease, polymerase and ligase. 5' exonuclease digests the 5' end of dsDNA...properties of type IIS restriction endonucleases. These endonucleases cut dsDNA at a specified distance...) relies on the 3'-5' exonuclease activity of T4 DNA polymerase. An exonuclease is an enzyme which removes...Restriction Enzyme Cloning Restriction enzyme (endonuclease) based molecular cloning is the "classic" cloning...naturally produced by certain bacteria and archaea, cleave double stranded DNA (dsDNA) at specific sequence...cloning project. Restriction enzyme cloning also leaves behind a short scar in the DNA sequence and can...cloning utilizes the Taq polymerase which naturally leaves a single adenosine (A) overhang on the 3' end of...

...targets may not result in equivalent cleavage efficiency. In fact, cleavage efficiency may increase or decrease...are denatured-renatured, treated with a nuclease that cleaves DNA heteroduplexes, and run on an agarose...decrease expression of a target gene? Once you have a clear understanding of your experimental goal, you are...Infects dividing and non-dividing cells AAV is the least toxic method for in vivo viral delivery RNA delivery... sequence and target DNA may result in reduced cleavage. The number of alleles for each gene may vary ...likely to result in the most efficient on-target cleavage. The gRNA target sequence needs to match the target...gRNA design. In general, off-target sites are not cleaved as efficiently when mismatches occur near the PAM...

...code inserted into the plasmid. This produces the least amount of variation between antibodies, allows for... induced to express antibodies. Antibodies are released into the growth medium and can either be used ...fragments - Fab fragments are created by enzymatically cleaving the variable regions of an antibody (each Fab ...they should be stored at -20 °C in aliquots of at least 10 µL. Antibodies cannot tolerate freeze-thaw cycles...single cells. Includes: Flow cytometry Read on to learn more about the applications common to each category...should be run with your protein of interest and at least one ubiquitously expressed protein such as GAPDH...protein:DNA complexes. After capture, the DNA is released from the complex and can be sequenced and amplified...

...Ready to learn about antibodies? Our latest animation is here to help! Join Abi as they explain what ...

... to take our word for it!  “When people want to learn about plasmids, they don’t go to Wikipedia; they...

...fundamental technical lab skills, like pipetting! We’ll release a new tip every two weeks during the school year...

...second edition includes new and updated content in a clean and easy-to-navigate design. You can find it, along...

... "what is an IgE isotype?"  @addgene Ready to learn about antibodies? Our new antibodies animation has...

...burning AAV questions answered.  Interested? You can learn more here - and remember, submissions are due Nov...

...here Additional resources on the Addgene blog Learn more about Blugene's make-over Get a how-to guide...

...BioRender.com. Figure 1: PRIME editing Please check out the updated guide for a great introduction...

...episode of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for...

...episode of our Hot Plasmids podcast series, you'll learn about new libraries for studying B. subtilis biology...

...programmed genome editing by a miniature CRISPR-Cas12f nuclease. On the eighth day of CRISPR, Addgene gave to ...

... nonprofit. Recent remarks and actions by X’s leadership conflict with our values. For this reason, our...

...photos of our pets with Addgene gear, and have unleashed them all over our recently redesigned blog. 🐕...