...insert targeted mutations through homologous recombination. To further increase the utility of CRISPR/Cas9...containing the RNA pol III promoter, gRNA and pol III terminator. These relatively large cassettes (considering...Size is also a factor; the pol lll promoter and terminator need be encoded only once, and each <200 bp tRNA-gRNA...

...ligation assay (PLA). The clever combination of FUNCAT and PLA PLA was originally published by the Söderberg...16590564. PubMed Central PMCID: PMC222790. 6. Buxbaum, Adina R., Bin Wu, and Robert H. Singer. "Single β-actin...

...questions the conclusion of inter-homologue recombination was released. This blog has not been updated...al. were interested in correcting an autosomal dominant mutation in MYBPC3 that leads to hypertrophic ...embryos to find those that do not carry a given dominant disease-causing gene, or that carry only one copy...

...time). Other illumination strategies, such as 2-photon-excitation, selective plane illumination, TIRF or ...experiments on a single day on a stable system. A final piece of advice is to measure the excitation power...

...yellow, or magenta, depending on the hairpin combination bringing the total number of individual colors...Adding an additional hairpin/fluorescent protein combination, for example, with a far-red FP, would increase...15! They also envision using CRISPRainbow in combination with gene editing. CRISPRainbow requires very...

...available (below). However, on November 22nd a preliminary injunction against the FLSA updates was granted...inconsistencies across geographical regions When we originally started gathering this information, many assumed...to pursue academic work, and to enhance the sustainability of the scientific enterprise by reducing the...

...or awaiting a final publication in a journal.  Preprints can be one tool in a combination of strategies... Addgene ID for their deposited plasmids in the final journal publication or an updated preprint, further...

...played enormous roles in the development and dissemination of fluorescent protein tools. This year, we’...creates a painful sensation. The dorsal horn of the spinal cord is the first site of sensory input from our... the dual virus expression in CR neurons of the spinal cord. Created with BioRender.com. Image from Haoyi...

...This post was originally written by Jennifer Tsang and updated by Rachel Leeson. Have you ever wondered...cloning, waiting, and repeating before you can finally dive deep into generating the data you’re excited...cost $155.25 in reagents and consumables today. Finally, this doesn’t account for the most valuable component...

... deletions.   Cas12a – The staggered cutter  Originally classified as Cpf1, Cas12’s primary difference...Another similarity to Cas12a is that Cas14 also indiscriminately cleaves non-complementary ssDNA when engaged... the genome. Cas13, similar to Cas14, also indiscriminately cleaves ssRNA when engaged at its RNA target...

...drug discovery process.” The ClonTracer library originally came about as Bhang and co-workers were trying...treatment process. The authors went on to show that combination treatment with multiple drugs reveals even smaller...heterogeneity to drug discovery and the development of combination therapies.         References 1. Hyo-eun, C. ...

...is ideal because it eliminates the potential of human error or cross contamination between samples. With... Shannon Rinaldi...

...immunogenic profile. AAV was first discovered as a contaminant of adenovirus preparations. It is an icosahedral...untreatable until 2008. Following a single dose of subretinal administration of rAAV2 carrying the human gene... responses to adeno-associated virus and its recombinant vectors." Gene therapy 10.11 (2003): 964-976....

... the protein or structure of interest. For a C-terminal tag, as shown in the example above, we inserted...would be transcribed and translated. For an N-terminally tagged protein, we used the same strategy, inserting...tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization." bioRxiv (2017): 123042. Additional...

...dCas9. Within a few rounds of cell division, the original methylated strand is diluted to undetectable amounts...dCas9 so that it may later be excised by Cre recombinase. Addgene has several great options: pLV hUbC-...L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function...

...Originally published Apr 30, 2020 and last updated Jul 12, 2021. What started out as a small collection...used for this study at Addgene which include a C-terminal deletion in the spike protein to improve expression...pseudoviruses to determine how effective sera from vaccinated individuals are at neutralizing the different...

... Editing, and that meant rereading many of the original papers reporting various prime editing tools. ...your curiosity, we encourage you to check out the original papers for more details. You can find many software...prime editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652...

...AAV serotypes, and the ever-growing variety of recombinase systems. Picking Ligands Picking the right ligand...ligand co-developed by Bryan Roth's and Takafumi Minamimoto's lab called deschloroclozapine (DCZ) shows promise...our chemogenetics poster! This blog post was originally written by Maya Peters Kostman in May 2020 and...

... incident, and the sterilization of women in marginalized communities have created a legacy of distrust...included taking extra coursework and attending seminars in bioethics, public health, science policy, and...advantage of their academic environment and attend seminars, symposia, or courses in social, health or policy-related...

...majority of our plasmids and pooled libraries were originally described in a peer-reviewed scientific publication... our thousands of depositors. That means their original articles are your best bet to figure out how to...asking to avoid unrealistic expectations. And finally, provide feedback. If you are unsatisfied with ...