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...introduces himself and explains why he decided to do an MD PhD - it's all in the family. 3:56 - 7:04 :...gene drives work and what work he's planning on doing with CRISPR-based gene drives focusing on safety...

...Duedu inspire new Ghanaian scientists to continue to do excellent work in the biological sciences. Future...plasmids from the Addgene community and hope we can do even more to enable scientists in developing regions...

... model a specific disease or process of interest. Do you want to: Generate complete and permanent loss...target cells are not transcribing DNA to mRNA, nor do they need to translate mRNA to protein. Instead, ...a lack of efficient target cleavage in cells that do express both Cas9 and gRNA. Before choosing a validation...using cationic lipids, electroporation, or an endosomal approach. Similar to the mRNA method described...

...they do in the software. Most sgRNA design tools require the user to specify an organism. They do this...what the parameters represent. The best learn-by-doing tool is E-CRISP. The authors have made a huge effort...for labs: Define what you need the software tool to do before going shopping for one. Use the CRISPR Software...criteria. Repeat until you have found the best tool. Do several experiments and use the results to re-evaluate...make a difference in our experiments and lives. To do this effectively, there needs to be good communication...

... to top Content last reviewed on 01 October 2025. Do you have suggestions for other plasmids that should...

... cost. The problem with these assays is that they do not provide sequence-level data. They also have a... a limit of detection of ~5%. This means they do not reliably detect editing events that occur in less...variable) Sequence-level data, extremely sensitive Do not sequence all DSBs, may miss unpredicted off-target...Tsai et al. 2015). Additionally, current algorithms do not take into account other elements, including those...GUIDE-seq and BLESS detect double-stranded breaks and do not require high sequencing read counts making them...

...take to reduce them. But we do know we will continue to ask “What can we do to make our materials both ...by researchers all over the world — and we really do mean all over (Figure 3). We continue to add new ...more reproducible science and learn what Addgene can do to help. More resources on the Addgene blog Twenty...

... was doing the kind of work my lab wanted to do already seemed to know each other and what to do.  Saved...tracks), something we didn’t have enough people to do in the pilot. We also found that all participants...

...video background! (720 px or 1080 px).         Do you do a lot of transformations with the plasmids you...

...Golden Gate etc.), and try to find the simplest way to do it (i.e. avoid assembling too many fragments at once...by methylation like XbaI, ClaI, etc. If you are doing multiple digestion be sure that the buffers and ...plasmid, and analyze it using the following steps: Do a single or double digest to be sure that the plasmid...

...need to sort through them to find the desired clone. Do your selection, then screen Why add a screen to your...plasmids that contain the gRNA (white) from those that do not (blue). Let’s take a look at a few screening ... containing plasmids with inserts from those that do not is by using a restriction digest. The key is ...

...phenotype in the surviving population. Selections do require there to be a culture condition that can ...sound like a great shortcut, right? Who would ever do a screen when a selection can get you to the answer...culture condition which eliminates manipulations that do not affect the function of interest. An example where...culture cells in the drug of interest at a lethal dose for non-manipulated cells and then assay cells which...

... synthetic modifications, specifically ones which do not contain a 5’ triphosphate in the gRNA, reduce...many suitable dyes to choose from and you can opt to do-it-yourself with a kit or purchase it commercially...but you can still choose what dye to use  What to do once you’ve chosen a gRNA and modification(s) Congratulations...

...we understand what genes do! Viruses have several options to allow you to do exactly that: shRNAs are...clinical promise moving forward. What viruses can do for you Whether you are interested in basic science...long-lasting immune response even after a single dose. Vaccine development strategies. Image courtesy...

...inducible? How long do you need control? Will temporary control (a few days) be enough, or do you want a permanent...101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose for my Experiment Additional resources ...

...groups and individuals with similar ideas to actually do something about it. Building a diverse team with ...There are many brilliant minds out there, but how do you get them to connect? Through networking of course...solving a community’s issue without involving them can do more harm than good. Check out this webinar for more...

...previously identified Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the...here Find the Konermann et al. Plasmids here How do Cas13d enzymes compare to other Cas13s? Like other... and is thus HEPN-independent. These enzymes also do not require a protospacer flanking sequence, so you...

...” of mentors as one key to their success. But how do you find these elusive teachers, supporters and advisors...be read or watched during the meeting.  Scientists do better with process, so don’t just get together and...blog in this series will have more ideas on what to do once you are in a mentoring relationship.  Celebrate...

...double-stranded DNA (dsDNA) genome. Non-enveloped viruses do not have a lipid bilayer surrounding the viral particle...cells for rAdV vector production, as HEK293 cells do not contain E4. Download the Bert Vogelstein’s lab...infection, wild-type adenovirus as well as rAdV vectors do not integrate into the host genome. Instead, they...cell line. Recombinant adenoviral vectors typically do not integrate in the host genome and instead are ...for recombinant adenoviral vector production that do not contain GFP. Recombinant adenoviral vector (rAdV...

...Resource Guide Content last reviewed: 23 October 2025 Do you have suggestions for other plasmids that should...