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... genome engineering in bacteria, alongside more established tools such as Cas9-recombineering, CRISPRa...and bind target DNA sequences using RNA-DNA base pairing. Separately, TnsABC forms a transposase complex... two transposase proteins (TnsB and C). While requiring fewer components for RNA-guided DNA integration...).  CRISPR-transposons for bacterial genomes engineering In the Sternberg group’s initial report of VchINT.... For transposons to be specifically recognized during transposition, genetic cargos must be flanked by...currently studies CRISPR biology and CRISPR genome engineering technologies. Message or follow him on twitter...high-efficiency, multiplexed bacterial genome engineering. Nat Biotechnol. https://doi.org/10.1038/s41587...

...distribution is recovering from the pandemic-related slowdown. More scientists are sharing plasmids and ...are able... Thanks to everyone for being awesome during a tough time. We're all going to get through this...This is a time when we need as much open science sharing as we can get. The effects of Addgene’s brief...Addgene Team because they’re all doing amazing work during this difficult time. One function that rose to ...had to figure out a way to maintain our services during the crisis and maintain our workforce for when ...enough funds to make most of our payroll for the spring and early summer months. We are also extremely ...for grant-driven research being done in all labs sharing materials via Addgene. Perhaps if each of many ...

...without looking at what they’re delivering and where they’re delivering their payload to.   Figure...anniversary celebration, we’ve been finding and sharing stories unique to our repository and resources....prioritization happens for good reasons. We began offering our viral vector service because we realized that... are available quickly. Beyond the service This brings us to the final piece of the viral vectors service...our AAV Data Hub as a community platform for the sharing of that exact data. Here, requestors can upload...their experimental data in a standardized format, sharing how well a given viral vector worked — or didn’...save time in selecting experimental conditions. Sharing our experiences Addgene’s plasmid repository gave...

... a mug or reusable water bottle, you’d probably bring in a mug or reusable water bottle instead of buying...newsletter Help other scientists save resources by sharing plasmids Find plasmids at Addgene ...

...roles. Therefore, institutions are responsible for ensuring that either all affected scientists are paid above...here) pushing for exemptions for postdocs, and lowering of the proposed salary level for exemption. Discussions... are institutions doing? Institutions are now preparing to comply with the changes to the law on Dec 1st...of 340 institutions with postdocs in science, engineering and health from the NSF’s 2014 Graduate Student...University of Delaware has a minimum of $36,360, requiring an increase of 30.6%. We have attempted to check...

...removal helps prevent the invading genome from “recovering” its original sequence. If you’re looking to ... High Fidelty Cas9 Mutants Browse All Genome Engineering Posts Additional Resources on Addgene.org Check... Mary Gearing...

...base pairing, it has promise to be host-independent, beyond the oft published metabolic engineering realm...summarizes Zalatan JG, et al.'s recent paper, "Engineering Complex Synthetic Transcriptional Programs with...of research has shifted in recent years toward tinkering with regulatory networks and multiplex gene control... ‘biofactories’, most successes in metabolic engineering have been limited to the overexpression of various...scaffolds follows the rules of Watson-Crick base pairing, allowing for rational, straightforward design....scRNA design is readily apparent in metabolic engineering. The authors elegantly demonstrated the potential...violacein, a distinctive pigmented protein. By altering the transcriptional programs, the authors sequentially...

...talk, which focused on the history of genome engineering from ZFNs through TALENs and CRISPR-Cas nucleases...huge impact on this field. Advances in genome engineering & CRISPR technology That afternoon's workshop...workshop, entitled "Advances in Genome Engineering", brought us 8 short talks on every conceivable way of examining...more parts-oriented synthetic biology. I've been answering biology customer service questions for nearly ...scientists are tackling this type of genetic engineering is a testament to how amazing the downstream ...downstream payoff could be. Though I attended an engineering school as an undergraduate, I never fooled myself ... The talks we heard Monday night represented engineering first and foremost, from the custom software ...

...jwoodgett.     More Discussions About Data Sharing: PLOS Data Sharing Policy Discussion PLOS’ New Data Policy...this year when it announced its data access and sharing policy. Since early March, the open access publisher...reasonable. PLOS has since clarified their data sharing policy and recently announced that of the 16,000...funding agencies (e.g. Wellcome Trust, NIH Data Sharing Policies, etc.) and is founded on the principle...data should be included? We should recognize that storing data in a sharable form is indeed a significant...Publishers have also contributed to this issue by requiring ever more condensed methods descriptions, legends...existing, published data. The importance of data sharing The reason this topic is important to my lab is...

... developed during the COVID-19 pandemic. Our mission to accelerate research through sharing reagents has...2021.03.10.434840 Tsang J, LaManna CM (2020) Open Sharing During COVID-19: CRISPR-Based Detection Tools. The...detect SARS-CoV-2 nucleic acids. Mammalian targets During the 2003 SARS outbreak, Hyeryun Choe’s lab identified...-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their search, they found mucins...inhibitors of SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV-2 viral...academic and industry scientists through plasmid sharing and more. As you’ve read above, some of these plasmids....     References and resources References Biering SB,  Sarnik SA, Wang E, Zengel JR, Sathyan V, Nguyenla...

... learning. 1. Create a peer mentoring group We don’t all get the mentoring support we need, and most of...Own Peer Mentoring Group: A How-To Guide for Scientists.   Once in-person, peer-mentoring groups ...use more mentors to listen, guide and support us during training and throughout our careers. An easy way...goals.  I’ve been surprised to see that virtual mentoring groups seem to be as successful as in person groups...? You could do a talk virtually for your peer mentoring group (see above). However, now that we are all...present, I’m asked what Addgene “looks for” when hiring a scientist. Of course, we have different criteria...

... take a moment to review what exactly you are measuring and how to interpret your results.  Quick note...Purification reagent contaminants The reagents used during nucleic acid purification can leave behind a variety... more considerations and other approaches for measuring DNA quantity and quality than we have covered ...Resources on Addgene.org DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol ...

...and its use as a genetic engineering tool is frequently called recombineering - short for homologous recombination-mediated...components of the Lambda Red recombineering system The lambda red recombineering system has three components...for generating dsDNA recombineering substrates to avoid errors introduced during PCR. ssDNA Substrate ...substrates can be generated by ordering synthetic oligonucleotides or denaturing short PCR products. Either..., Court DL. Recombineering: A Homologous Recombination-Based Method of Genetic Engineering. Nature protocols...Costantino, N. and Court, D. L. 2014. Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination...method that can be used for cloning or genome engineering and is based on homologous recombination. It ...

...interference, these enzymes can knock down RNA without altering the genome, but Cas13s have higher on-target specificity...enzymes are the latest tools in the transcriptome engineering toolbox. How were Cas13d enzymes discovered?...MAPT, the gene encoding dementia-associated tau, delivering dCasRx and a 3-spacer array targeting the MAPT... tell how large the genome and transcriptome engineering toolbox will be. It is, however, certain that...that the impact of CRISPR scientific sharing will continue to grow, and we at Addgene appreciate our depositors...References Konermann, Silvana, et al. “Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors...  Mary Gearing...

...by Benoit Giquel. CRISPR has taken the genome engineering world by storm owing to its ease of use and utility...Waltz, 2016), the potential of CRISPR for genome engineering in plants is also being realized. There are a...approaches, these techniques are cumbersome, often requiring several rounds of selection to isolate plants ...plants with the phenotype of interest. Genome engineering, on the other hand, allows for targeted modification...will present a general overview of plant genome engineering using CRISPR, highlight the specific modifications...plants, and outline the various plant genome engineering tools that are available to academic researchers... Addgene. CRISPR components for plant genome engineering CRISPR can be used to knockout, activate or repress...

...Collection under the insect expression system or by filtering by fly as species of gene. Find Gal4 and UAS plasmids...select for transformed bacteria, but you might be wondering, “How do I know which of my flies have the desired...about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific Recombination... genome engineering...

...important role in scientific exchange. In addition to ensuring affordable access to materials, repositories provide...since 1996.       More Blog Posts about Scientific Sharing: The 10 Most Distributed Plasmid Technologies ...10 Years Data Freedom: The Expansion of Data Sharing in Research Publications ...

...contributions from depositors around the globe, we’re now sharing over 100,000 reagents including plasmids, ready-to-use...

... to academic scientists. Expanding plasmid sharing during COVID-19 Realizing this limitation, we worked...members of the scientific community support open sharing. But, one critical part was missing when the COVID...setting.  In light of the need for basic research during the current pandemic, universities are showing ...cells. We teamed up with Ginkgo Bioworks to quickly bring these plasmids to the scientific community - both...available to industry scientists To make plasmid sharing more open for academic and industry scientists,...the past months, we’re excited to see our open sharing impact grow beyond COVID-19. But, we can’t do this...office to discuss the possibilities for expanded sharing. Is there a plasmid you're hoping will be come ...