...sharable products. Open-Source Microscopy Sharing Hardware vs. Sharing Software In the field of microscopy,...relatively young movement of sharing open hardware has a similar goal. The sharing of modules, ideas, and blueprints...material (BOM) of an assembly, the feasibility of manufacturing, and subsequent assembly can be difficult to... or 3D printers are key to enabling this work. Sharing open-source plans that rely on such tools has allowed...3D printing, on-demand CNC milling, and PCB manufacturing. These newly developed techniques are the key...enablers for making assemblies open-source. The sharing of digital representations of the 3D models, which...especially with regard to licensing models. Direct sharing of work from universities and research centers ...

... efforts check out Leila Haery’s blog series, “Bringing Sustainable Practices to the Lab.”  Whether participating...newsletter Help other scientists save resources by sharing plasmids Find plasmids at Addgene ...

...worry a very little bit about hiring when we need to do it quickly. Hiring is hard but as a 3 year Boston... space here at our new location to grow. We are hiring again and getting ready to embark on new exciting...distributed in 2018 and we are poised for more offerings. The viral vector team is publishing research,...We must keep spreading the word about materials sharing, but we don’t have to worry about getting support...False, type='scriptV4', hide_playlist=True, viral_sharing=False, embed_button=False, width='1280', height...

... too. Schedule regular meetings Scheduling a recurring meeting every two weeks or every month is long...  Maintain informal communication In between recurring meetings, you should share interesting results...they’re set by checking in about them at your recurring meetings and reaching out if any milestones are...

...resources. We've got twenty years of experience we're sharing through blogs, eBooks, videos, guides, protocols...

...terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration, producing a second copy...voltage fluctuations, while having high fidelity during action potentials of fast-spiking neurons. The ...baseline voltage), the investigators found that administering these AAVs with a dilute Cre-expressing vector...splitting a large gene into smaller fragments, delivering them by individual AAVs, and then splicing their...

... relationship to signal depend on what you’re comparing your sample to. For example, we use our negative...screening and development of a focused reference bring us one step closer to resolving the antibody crisis...

...circuit by integrating neuronal signalling and triggering release of the stress hormone norepinephrine ...Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent IL-1 beta overexpression cassette...lit (risky) and a dark (protected) area. When measuring the time spent in a particular part of the apparatus...levels within the hippocampus and frontal cortex during depression, yet there have also been contradictory...

...SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR)...

...waste. This is the second in a series of posts on bringing sustainable practices in the lab. The first post...waste from the landfill, but what about all the manufacturing steps that happen before you even receive the... lower the environmental impact in terms of manufacturing, energy, and water use. ACT labels are obtained...newsletter Help other scientists save resources by sharing plasmids Find plasmids at Addgene ...

...articles in this series: Bringing Sustainable Practices to the Lab: Innovations Bringing Sustainable Practices...Labconscious. I was eager to learn about practices I could bring both to Addgene, and share with my peers. In fact...the rate at which that happens and reduce the manufacturing of new containers, which is associated with ...huge contributor to the increase in plastic manufacturing, accounting for 40% of total plastic usage (...to be sustainability leaders. Take initiative to bring ideas to your workplace and you could end up making...newsletter Help other scientists save resources by sharing plasmids Find plasmids at Addgene ...

...Yoshii, Saori R., and Noboru Mizushima. "Monitoring and measuring autophagy." International journal of molecular...mitochondria-targeting sequence makes Keima useful for monitoring mitophagy. It’s important to remember that because...3B, or LC3. LC3 is a commonly used marker for measuring autophagic flux. mRFP-GFP-LC3 functions as an ...Maria Cuervo, and Per O. Seglen. "Methods for monitoring autophagy from yeast to human." Autophagy 3.3...

...imagination of most of the authors during this period.  Genetic Engineering Era: Pre-CRISPR  Having visualized...associated with genetic engineering in the recent SciFi movies. Rampage (2018), starring Dwayne “The Rock” ...California, San Diego. The revolution in genetic engineering techniques is a speculation of yesteryear which...but the common themes that these authors were exploring were space and robots. Warping to stars in space...devise methods to manipulate it. The term genetic engineering itself is attributed to SciFi writer Jack Williamson...s Island. In 1976, the world’s first genetic engineering company, Genentech, was founded.  This commercialization...these corporations would extrapolate genetic engineering to exploit human beings. Ridley Scott directed...

...preparation steps of production, purification, and titering. Our newest viral vector service, AAV Packaged...

...Addgene blog Twenty Years of Addgene Sharing: CRISPRTwenty Years of Sharing: Addgene's Viral Vector Service...many countries, import and export regulations, differing customs processes, and the distances the packages...specific to that country. ‌For example, when we began offering temperature-sensitive materials, we noticed a ...shipments arrived in India at room temperature. Partnering with our distributor allowed us to ensure that...shipments weren’t able to clear import control. Partnering with a distributor allowed us to share materials...scientists in China. They were also able to help with sharing information through the use of popular Chinese ...solutions aren’t enough to ensure accessibility, partnering with distributors who have expertise in a specific...

...chromosomal DNA.  Plasmids are said to be under stringent control when they depend on the host cell’s initiation...consider the replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...replication for the bacterial chromosome and for stringently controlled plasmids. Binding of IHF (green) redistributes... cell growth or even kill the host cell. For stringently controlled plasmids, replication is tightly coupled...number.  Let’s consider pSC101. This plasmid is stringently controlled and has a low copy number. Like the... necessary. Iterons are not the only tool to stringently regulate plasmid replication. For example, the...Comparison of replication regulation mechanisms of stringently controlled plasmids pSC101 and ColIb-P9. pSC101...

...2012). This made them an appealing target for bioengineering, but it wasn’t clear whether they could 1) ...bodies We have shown that all three of these engineering objectives are possible in our recent paper (...from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer, K.-H., Verni, F. & Rosati, G. Defensive...

...cycle for titering as this will better represent future experiments. The aforementioned titering options...in the long run. Read on for an overview of the titering options as well as the benefits and drawbacks ...measurements via ELISA and qPCR for viral RNA. Measuring p24 levels in a viral prep is straightforward ...transfer plasmid was used in the transfection; if preparing several different lentiviruses in parallel, this...should be considered. Similar to the p24 assay, titering via measurement of viral RNA can overestimate ...viral particles. One of the si mplest methods of measuring functional titer uses FACS to count the number...antibody staining. Check Out Our In-Depth Fluoresence Titering Protocol Another popular approach utilizes the...

...genome engineering have played a huge role. For instance, researchers are genetically altering tumor cells... works on genome engineering and cancer immunotherapy, and is interested in exploring new avenues in healthcare...ve been studying it and, now, researchers are exploring ways to bolster and modulate our immune defenses...environments for tumor cells spreading to other organs during metastasis. Evading Recognition Tumor cells over-express...cause a transient shutdown of the immune system, hampering its ability to inhibit further cancer growth and...increasing life expectancy by 4.1 months as opposed to curing the cancer. DCs are harvested from the body, and...trials going amiss and patient deaths have occurred during CAR-T cell tests because treatment regimens were...

...also a story of community, cooperation, and sharing. Sharing this story is a truly special way to celebrate...As we celebrate twenty years at Addgene, we are sharing stories of our repository’s positive impact on ... that developed CRISPR tools were committed to sharing them with the rest of the community. They did this...the usefulness of popular CRISPR tools is both enduring and rapidly recognized.   Where is the CRISPR ...appropriate MTAs and customs forms. Of course, ordering the plasmids is the easy part; using them can ... resources on the Addgene blog Twenty Years of Sharing Science  ...

...of modified Ti plasmids during plant infection, as well as plasmid yield during preparation in E. coli....the plants that we grow and eat, and genetic engineering will continue to improve plant nutrition, yield..., and pest resistance. Much of plant genetic engineering revolves around Agrobacterium tumifaciens. Agrobacterium...Agrobacterium to ensure that they are maintained during co-cultivation with the target plant species. Thus...with biolistic methods, making it useful for engineering plants that are not susceptible to Agrobacterium...customizability, pSiM24 can be used not only for plant engineering and biotechnology applications, but also for ... plant genetic engineering...

...these defenses. For some applications of genome engineering - such as certain forms of gene therapy - avoiding...tool and an alternative to CRISPRs for genome engineering whose use is only likely to grow in the coming...  genome engineering...

...stably integrates into the genome and is propagated during normal DNA replication and cell division. Mobile-CRISPRi... these plasmids for transfer to recipient cells during a tri-parental mating scheme. Mobile-CRISPRi integrates...was variable. They measured transfer rates by comparing the number of colonies that grew on a selective...about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi, and CRISPRa...

...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

...-2, but with many academic research labs closed during the pandemic, it’s hard to get these reagents. ...available through a repository, they remain accessible during these times. Centralized reagent repositories, ... rapid access to validated reagents, especially during a time when these materials may not be available...for further investigation of this pathogen. The sharing and distribution of cell lines from repositories...community will continue having access to plasmids during the COVID-19 pandemic. Over 7,000 plasmids for ... Case study 3: Mouse strains Reagents developed during the SARS-CoV pandemic more than a decade ago are...et al., 2006). Now that a similar phenomena is appearing with regards to SARS-CoV-2, this mouse strain ...

... course the rapidly changing world of genome engineering via CRISPR.  Though the individual topics are...

...been able to progress so rapidly is scientific sharing. Already we’ve seen articles about COVID-19 on ...centralized databases and centers to support the sharing of materials and data associated with COVID-19 ... these resources and why they value scientific sharing as the scientific community strives to understand... motivations vary, everyone we spoke with sees sharing and collaboration as the best way that the scientific...are supporting their access to useful resources during this time. Facilitate collaborations within ...researcher studying COVID-19. “Open scientific sharing is important for creating research synergies and...iterating on methods for coronavirus research, sharing resources for COVID-19 research in their field ...

...determine functional titer with a colony formation titering assay or similar protocol. After functional titer...skewed the representation of your pooled library during the amplification process. It’s far better to verify...worked.  Ready, optimize, transduce!  If you’re ordering a pre-made library, you don’t need to worry about...optimize before you can transduce your cells.  Titering After generating your lentivirus, you’ll want ...specifically to allow scientists to perform functional titering and optimize transduction efficiency. Transduction...page from our book and put aside an aliquot for titering and optimizing before you freeze down your viral...and validation of CRISPR libraries. The Genome Engineering using CRISPR/Cas Systems Forum- Moderated by ...

...included continuous outflow of phage, increasing stringency and promoting survival of only the broadest-targeting... Mary Gearing...

...CRISPR Plasmids Find More Tools for Combinatorial Engineering on Our SynBio Pages ...

...for positions both inside and outside academia. Inspiring future scientists At Addgene, we’re incredibly... a scientist and what benefits did they provide during your education? Let us know in the comments section...

...experience working on and later chairing the Advocacy Committee. I’m now furthering my leadership experience ...abilities we often hear that scientists should acquire during their PhD and postdoctoral training. Addgene executive...more substantial experience can be achieved by volunteering for a professional organization or a scientific...Vision and Ophthalmology Research (ARVO). Hopefully sharing my experience will show you how you can get started...started on your own development as a leader. Chairing a committee I was Chair of the Advocacy Committee...updated some sections of the NPA’s website. By volunteering, you can gain experience working with other ... my verbal and written communication skills. Volunteering on these committees was real work; meetings ...

... start a peer mentoring group? "What Makes a Good Mentor?" Form Your Own Peer Mentoring Group: A How-...presentation skills (as much as you can), form a peer mentoring group to get real feedback Write whenever you ...How-To Guide for Scientists Download our eBook: Mentoring for Scientists [PDF] ...

...about her mentoring style. Set goals with a mentor: The secret of accountability To make a mentoring relationship...want more great advice on mentoring, click below to download the "Mentoring for Scientists" ebook, which... the fifth and final post in the Addgene Blog Mentoring for Scientists Series. The entire series and additional... If you have been following the posts in this Mentoring for Scientists series, you have: realized the ...adding formality and active goal setting to your mentoring relationships, so that you can reap rewards in...Joanne's Reddit AMA Add a little formality to mentoring Just getting together and chatting with another...about their career path is a great start to a mentoring relationship, but if you don’t take this past ...

...brain at a level of abstraction that enables the engineering of its function." Once scientists can do that...big questions that motivate his interest in neuroengineering, and some of the optogenetic tools that are...are still missing. Optogenetic tools for neuroengineering Addgene: For those who may not be experts, can...set of neurons, maybe we could treat patients suffering from Parkinson's, chronic pain, deafness, or other... neuroengineering...

...article in the Addgene Blog Mentoring series will cover 2 of the 7 mentoring questions I set out to answer...programs A formal mentoring program can be a great way to get started with mentoring. Read this Catalyst...Catalyst Report “Making Mentoring Work” to learn about what makes a good mentoring program. If your organization...also a great place to practice your mentoring skills. Online mentoring can work if both parties are diligent...my upcoming mentoring post in which I describe how to organize a successful peer mentoring circle. Asking...School researcher Connie Cepko to learn about her mentoring style. Mentors at home and away We should all ...organization has a mentoring program, consider tapping into it and helping make it work by being an educated...

... 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 ...

...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...

...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...

...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...

... 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...

...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...

... 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...

... 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...

...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...