...disease research and provide hope to patients suffering from these devastating afflictions. Elia Di Schiavi...for peritoneal mesothelioma and thereby relieve suffering from this little-known disease. You can find more...Nabavi and will continue to work to help those suffering from rare diseases. These underrepresented individuals...

...co-op) is what has brought me, a second year bioengineering student, to Addgene. I am currently working...Addgene, I felt comfortable asking questions without fearing they would be disregarded - everyone was very approachable...comfortable.     Lucie Wilson is a rising junior and bioengineering student at Northwestern University. She is ...

...preferences using HEK293 cells. They started by comparing D10A and H840A total editing efficiency in PAM-in...H840A creates 3’ overhangs in a PAM-out design.  Exploring nickases for homology directed repair The potential... Mary Gearing...

..., had to keep him from running too much as any jarring of his body could initiate very painful FOP flare...donation via the link below. Please continue to help bring HOPE to AJ.   PS: The superhero race is the perfect...running on behalf of AJ Gonzales, a young boy suffering from FOP, raising money for NORD for the past ...

.... If you decide not to cleave the GST tag after/during purification, be aware that the tag dimerizes in...purifications due to the presence of more naturally occurring histidine-rich proteins in mammals. Mammalian ...possible but requires optimization and often more stringent washing. Nonetheless, this tag can be purified...

...for 2-photon imaging of voltages. This includes monitoring of fast bursts, as well as slow up/down voltage...work also proposes promising avenues for future engineering to shift the spectra and improve the performance...not just because of the title!) The Myc-tag is a string of 10 amino acids, EQKLISEEDL, that can be incorporated...

...obstacles that stand in the way of the doers and by ensuring good communication. A manager must both hire and...delegating tasks and in motivating their doers by ensuring they are as engaged as possible. To learn more...of leader, one needs to take the initiative by bringing and effecting solutions to both known and unidentified...

...associated genes) system. This popular genome engineering tool offers flexibility, multiplexibility, and...system that scientists have harnessed for genome engineering. The diversity of the CRISPR/Cas systems provides...who has the strongest will to survive. Genome engineering and future directions CRISPR/Cas inspired genome...

...screens require a low multiplicity of infection (MOI) during cell library construction to ensure that the majority...screening, calcineurin dynamics measurements, or monitoring in vivo. To overcome these technical issues, ... proteins can bind specific DNA sequences which brings the attached fluorescent protein to the DNA. They...

...reached out to MIT labs in the Synthetic Biology Engineering Research Consortium. That is how Aditya Kunjapur...robust tools that have practical applications in monitoring the accidental or intentional release of engineered...complementary tools in a time where biological engineering and synthetic biology is expanding rapidly.  ...

...Agreements, your TTO signs each agreement just once, covering nearly all future requests/deposits. This reduces...Institute  California Institute of Technology Cold Spring Harbor Laboratory  Dana-Farber Cancer Institute...Hutchinson Cancer Research Center Memorial Sloan-Kettering Cancer Center  Monash University  Montana State...

...different CellTag libraries at multiple timepoints during reprogramming. These library-specific identifiers...different CellTag libraries at three time points during the 28 day study: prior to reprogramming (CellTagMEF...CellTagging. By constructing lineage trees and comparing them with the gene expression changes of more ...

...preventing them from moving forward? Answering these questions will bring you and your audience - via the ...sense of curiosity and wonder why you’re doing and sharing science in the first place?   References and Resources...

...pR6K ~15-20 R6K* C Stringent pACYC ~10 p15A B Relaxed pSC101 ~5 pSC101 C Stringent pBluescript ~300...of replication is referred to as "relaxed" or "stringent" depending on whether the ori is positively regulated...

...syndrome, a rare disease that can cause hearing loss and changes in coloring of the hair, skin, and eyes. The...competition for researchers working on rare diseases, offering them the chance to win some of the latest tools...

...considerable number of researchers, I had high hopes of securing tenure in a British or Irish University where ...how to sell these products to other scientists. Tailoring my CV for the job To become an attractive candidate...removed my techniques and protocols, courses taken during my PhD and Undergraduate sections and brought in...

...identify potential career pathways, education and mastering new skill sets must occur by doing work outside...can: Introduce new perspectives in science and engineering Looking at your research from a broader viewpoint...Networking As the number of qualified scientists entering the workforce increases, EAs allow graduate students...

...process. Even though this process is essential to sharing plasmids around the globe, it can be a bit of a...clear expectations from the beginning makes the sharing process smoother, more consistent, and more pleasant...MTA YouTube video for more information. Happy transferring!    Many thanks to our Tech Transfer team members...

...common ways to quantify DNA. This method involves measuring the absorbance/transmission of light through a...fluorometer to quantify DNA this way. First, start by pouring your gel containing a DNA intercalating dye (ex... the run time is just a few minutes per sample. During the run, DNA fragments move through a micro- or...

... and understand. All of this communication and sharing fits together to make for a transparent and collaborative... for data management plans and for evidence of sharing for grant renewal—but there is clearly room to ... all its different forms as part of graduation, hiring, funding, and promotion processes, I’m confident...

...pathways can modify the sequence of a gene (thus altering its function). HDR is crucial for certain applications...Moreover, the cost of reagents is relatively modest, requiring expressed Cas9 protein, transcribed sgRNA, and...al. "Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery....

...hypothesized that she could make enzymes controllable by tethering them to Dronpa, a fluorescent protein known to... HMOs are thought to play an important role in nuturing the development of beneficial bacteria in the ...HMO biosensors and displayed her findings at the Spring 2017 ACS meeting. She hopes these biosensors and...

.... While clearly an outlier, this story had us wondering: is there a larger trend here? Do papers associated...Management. She is particularly interested in exploring how innovative tools influence the direction of...References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." Science 339.6121 (...

...proteins or cells while having little effect on neighboring cells. The photosensitizer used in this work ...effective killing of dL5** cells without death of neighboring bystander wild-type cells.   Zebrafish experiments...photosensitizers had similar rates of off-target damage to neighboring cells. TL;DR MG-2I-dL5** is a NIR photosensitizer...

...the Wyss Institute for Biologically Inspired Engineering and member of the Addgene Advisory board, on ...aim to alleviate this frustration by producing, titering, testing, and providing lentiviral preps of select...the Wyss Institute for Biologically Inspired Engineering and the Zhang and Doench/Root labs from the Broad...

...from Janelia (jGCaMP7) continue to be useful for monitoring neuronal activity in the form of calcium dynamics...fluorescence and resting fluorescence (achieved through lowering sensor resting fluorescence, while maintaining...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

...serotype for their particular study, which includes comparing the transduction patterns of different AAV serotypes...its calcium-bound state, which was achieved by lowering the basal fluorescence of the sensor. This “contrast...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

...the weaponized T-cells (CAR-T cells) - and then delivering them to a tumor - they used focused ultrasound...staining, and light exposure can negatively affect FPs during these processes. The Liu and Petsko labs recently...only brilliantly bright, but it also withstands denaturing conditions, high temperatures, and other chaotropic...

...resulting from the catalysis of a chemical reaction requiring luciferin, ATP and oxygen as substrates. Production... have been identified in various species with differing kinetics, substrate requirements and photon emission...these luciferases have been subjected to further engineering to improve upon the properties of the native ...

...ends to insert genes into plasmid backbones, base pairing between these ~4 base overhangs isn’t strong enough... be accessible to allow for complementary base pairing, so SLIC can’t be used if the overhangs would have...ligase! This post was originally written by Mary Gearing in December 2015 and was updated by Emily P. Bentley...

...into our normal full plasmid NGS queue in order to bring them inline with our slower, more traditional, QC...from Ginkgo. To balance the need for speed while ensuring our rigorous QC standards are met, we are taking...’re here for and we’re glad to be able to do so during this critical time. Visit our COVID-19 Plasmids...

...these omissions, so it is unlikely to be caught during the peer review process, which often focuses more...respond as expected, use of representative images, measuring the main outcomes in several different ways, and... a particular result. Anything else that occurs during the course of the study, say a fifth time has unpredictably...

...until it is triggered to enter the lytic cycle.   During both of these life cycles bacterial DNA can be ...in a wild type E. coli (Griffiths et al., 2000). During conjugation, genetic material is transferred from...Guillemin. "Multiple phases of competence occur during the Helicobacter pylori growth cycle." FEMS microbiology...

...do not require breakdown of the nuclear envelope during cell division to enter the nucleus as gamma-retroviruses...AAV also has multiple serotypes (12 naturally occurring and more synthetic capsids are routinely being...lentivirus, retrovirus, adenovirus, and AAV. If you’re wondering more about what applications these viruses are...

...approaches can be found. As an example, say you are considering either a cellular screen or a selection. A selection...scene, there are other options which are worth considering. RNAi screens are similar to CRISPR, but instead...temperature sensitive screens. These involve raising or lowering the culture temperature to exacerbate a mutant...

...creates challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature CRISPR-Cas... Figure 4: Overview of CasMINI for genome engineering applications. Image from Xiaoshu, et. al. 2021...system opens up new possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular...

...terminator. These relatively large cassettes (considering the gRNA itself is ~100 bases) limit the number...examined editing efficiency in intact rice plants, comparing traditional gRNA constructs with PTGs. In PTG ... Mary Gearing...

...of experimental problems or questions you are pondering and be ready to talk about them. People love to... to help especially when solving problems.  During the conference The conference may start in the airport...post. Stay in touch by liking, commenting and re-sharing their posts. LinkedIn users notice this sort of...

...Sample6, for advice on how to take an idea developed during graduate school and turn it into a biotech company...changed, but Sample6 still leverages the core engineering and scientific concepts to engineer bacteriophages...co-founders raised $6 million. The biggest challenge during this time was learning to communicate effectively...

... really, and what new questions does this paper bring up? We’ll sift through the paper to understand what...results in fewer eggs. However, the potentially staggering costs of such a treatment on top of the expense... Mary Gearing...

...placing on your audience when you're in the midst of sharing your science - which can drive away even the most... them restate or summarize the concepts you’re covering. Don't show frustration if they're incorrect; ...as people discuss their experiences. If you’re hearing multiple people with similar misconceptions, it...

...polyclonal antibodies include any antibodies produced during the immune response.   Polyclonal antibodies...If your protein might become slightly denatured during your experimental protocol, using a polyclonal ... (2019) Antibody Purification and Storage. Cold Spring Harb Protoc 2019:pdb.top099101. https://doi.org...

...within both the synthetic biology and genome engineering fields. We’ll walk you through how to apply this... Engler & Marillonet. Golden Gate and genome engineering In early 2011, the Bogdanove and Voytas groups... Mary Gearing...

...mining tool, Transcriptomine. We are currently nearing the completion of an 18-month effort to overhaul...identifying plasmids in the research literature and acquiring them in a timely manner. Addgene represents an...an excellent solution to the problem of sharing plasmids, providing researchers with a one-stop shop for...

...extension begins with a C base can cause unwanted base pairing (left), while pegRNAs that use other bases in this...the cell to use the edited strand as a template during DNA repair. Test multiple nick sites, starting ... linkers that minimize unwanted intra-RNA base pairing with the primer binding site (Nelson et al., 2022...

...publications in the fields of research they support. Monitoring these types of accounts can help you find papers...papers later. Pro tip Learn more about authors by exploring author pages (for example, Melina Fan’s author... Instead, you create your own research feed by entering keywords, concepts, journals, and/or papers of...

...characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence triggers...and silences its expression. More specifically, during the process of RNAi, an enzyme dubbed Dicer cuts...smaller dsRNA fragments are referred to as small interfering RNAs (siRNA) and they bind to a class of Argonaute...

...out your samples. This can be useful if you’re transferring samples from 96 wells plates onto agar q-trays...bacterial samples at the same time as opposed to preparing each sample individually. Enter the BIOROBOT! ...brushes the information onto the vials instead of requiring us to put a label on each tube by hand. We’ll ...

...disparities. But as the science moves forward at a blistering pace, it is becoming ever more urgent for scientists...ways of collaborating, and to organize events that bring different voices together to talk about genetics...running these events will be important both in preparing you for future work in these areas, and in helping...

...create a successful science blog  5) Get creative – Bring your knacks into play Perhaps writing just isn’t...projects, you can set up a citizen science project. Answering the big science questions around climate change...guest blogger, Sarah Schmidt! Sarah Schmidt is engineering resistance against Fusarium wilt disease in bananas...