...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...

...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 ...

...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 ...

...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 ...

...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 ...