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...recombination-mediated genetic engineering (find more information on bacterial engineering using CRISPR on our blog..., their interactions and information processing during this condition is crucial. Plasmids expressing ...interactions. Combining synthetic biology with engineering: glowing bacteria and a fish robot to detect ...interesting combinations: biomedicine and astrobiology. During the last 11 years he has studied various aspects...

... HeLa cells?), genome engineering and stem cells, and the difficulty of sharing banked iPSCs across international...years, I've been looking forward to seeing how the sharing of his OSKM factors have affected the stem cell...paper as "life changing". Dr. Yamanaka himself, during his keynote address, appeared almost incredulous...into a pig. In stark contrast to recent genome engineering technologies, stem cell techniques are neither...

...for various scientific or engineering applications.   References Refactoring and optimization of light-switchable...photobiology: why use light? In synthetic biology and bioengineering, scientists seek to directly probe and build... publication at Addgene. Synthetic biology: engineering Escherichia coli to see light. Levskaya A, Chevalier... Mary Gearing...

...to the interview here! Doench majored in history during his undergraduate work at Hamilton College. When...that was open ended... it was actually something bordering on research, rather than 'here’s the protocol,... He first heard about RNAi at his MIT interview during a lecture given by Phil Sharp. After a few rotations... easier for people everywhere…. the nonprofit, sharing ethic that Addgene has behind it is something that... sharing...

...job. How can you incorporate the skills developed during graduate school into your resume? How can you stand...Here are a few things to consider when you're preparing your resume for a non-academic position: What ... current job as a Communications Coordinator. Wondering how you can develop more transferable skills? ...? See Developing Transferable Skills During Science Training and So You Think You have Skills.   Get Science...services, but some of the general knowledge about preparing a good resume, writing a concise cover letter,...

...off-target cleavage should be an important consideration during any experiment. Sensitivity Cas9 sensitivity is... the Cas9 nuclease. This can be determined by measuring the indel frequency at a target site. A specificity...not be accessible by wild-type Cas9, due to its stringent PAM specificity. This can be especially important...PMCID: PMC6368452. Strecker, Jonathan, et al. "Engineering of CRISPR-Cas12b for human genome editing." Nature...Repurposing CRISPR-Cas12b for mammalian genome engineering." Cell discovery 4.1 (2018): 63. PubMed PMID:...

...interest in sharing, there will always be an interest in protecting intellectual property. Figuring out that...of Katz and Paratore onto the Board of Directors brings complementary experience and insight to the now... to the scientific community through Addgene’s sharing platform. Expanding further beyond the US and increasing...Internal Audit function and is responsible for bringing a systematic and disciplined approach to evaluating...

...accessibility is removing barriers when we can and lowering them when we can’t. We know barriers exist in ...layers are blurred after centrifugation and the syringe needle is inserted bevel-up at the interface between...the 40% layer. To collect purified AAV, insert a syringe at the interface of the 40% layer containing the... iodixanol layer, with the bevel pointed up. A syringe at the top of the tube allows air to enter the ...the column so the virus will flow out the bottom syringe."Feel free to share your alt text in the comments...

...Before I started writing for the Addgene blog, sharing Chemistry Cat memes was how I used social media...professionally, and 2) I needed to start using Twitter. Sharing Chemistry Cat memes use to be how I used social...conference, whether you are attending in person or monitoring that conference via social media. The podcast...Twitter is a tool worth using, but you might be wondering how to get started. This blog post outlines 10...unfollow those who do not capture your interest during that week.” You can also find Dr. Knoepfler on ...

...components are still there in real life.  Certainly referring to any DNA within the context of a nucleus as ...methylation status at a double-strand break that needs repairing. They may be looking at chromatin at the level...! And if you study chromatin and want to start sharing your plasmids through Addgene, don’t hesitate to...Robert Kingston Thoru Pederson Phil Sharp Julie Ahringer Alexander Stark Bas van Steensel Richard Young...chromatin and the epigenome. If you have been considering some new techniques, the posts below may steer...

...” has me considering the microbiome more often than not). If you’re someone who’s considering getting ...Even if you're currently a synthetic biologist engineering E. coli metabolism, maybe you’ll be able to apply...because they provide a highly versatile means of delivering genes to many different cell types with minimal...Optogenetics, Neuroscience, and the Future of Neuroengineering Primer on Optogenetics Resources at Addgene.org...

...genome replication, repairing mismatched basepairs or small indels that occur during DNA synthesis, and...restriction enzymes are one half of naturally occurring restriction modification systems that prokaryotes...DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired mutation...methylation Finally, you can control methylation by altering your choice of bacteria. For example, if you must...

...most important step in ensuring quality, as it will catch any errors made during the cloning, sample handling...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...samples coming through our doors, the first step to ensuring plasmid quality is to stay organized. Here at ..., antibiotic resistance, restriction sites used during cloning, and most importantly, the predicted full...

...made of protein-coding genes, and you might be wondering what the rest of our genome could possibly be ... modulate ncRNA expression without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR...high specificity, particularly in applications requiring smaller delivery systems (Zibitt et al. 2021).... a number of different options. By carefully considering your ncRNA of interest, you should be able to...RNA-targeting CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer...

...completely different than what was originally intended. Sharing unverified plasmids can lead to far reaching consequences...standardization provides security for requestors, ensuring that the plasmid they ordered contains the correct...QC topics that may require additional attention during analysis. These include topics such as: locations... features. Most of these regions are identified during the QC process, following sequencing and assembly...30% of deposited plasmids! We are committed to ensuring that we maintain a high standard of sequence accuracy...

...retinal ganglion. Rather than using naturally occurring promoters to drive transgene expression, the Roska...one type of cell, with the remaining expression occurring in only Müller glia cells, a common contamination...retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic promoters performance in mouse, monkey...targeted cells in monkey and human retinas. By comparing the performance of promoters across these three...be paired for targeted expression. This same pairing of synthetic promoters were also used to target...

...their promises, Wet-blanket negativist, who can bring down the morale of a group with their pessimism,...communicate their ideas confidently while also considering the needs of other people. Some students and ...Regardless of your supervisor’s management style, bring challenges to his or her attention as soon as you...signed, or manuscripts that need to be reviewed, bring them to the meeting. This will make it easy for ...problem by stating the facts and explain how it is interfering with your work. Avoid talking about your emotions...

...special considerations. Conventional flow cytometry During conventional flow cytometry, light emitted by a...distinguish these two fluorophores from one another during spectral unmixing.     Figure 2: The ...a three color panel or a 40 color panel! When preparing a spectral flow experiment and panel, there are...telling exactly how many Cy7 molecules are added during that particular batch. This can present particular...upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical to include...

...something done not by people wearing a lab coat, but instead by people wearing business suits discussing ...transferable skills while working in the lab and by offering advice on areas where you might need some help...you are negotiating. Then take a moment before, during, or after a negotiation to think about the situation...experience in the area I want to explore while also ensuring that my work gets completed? I want my company...

...perform functional tests. The newest genome engineer sharing the stage with much-lauded CRISPR-Cas9 is DNA-guided... endonuclease NgAgo! We'll discuss how NgAgo is faring with users in a minute, but, to start, let's review...cleavage (1). Both the CRISPR-Cas9 and NgAgo genome engineering methods have their own pros and cons (2). The...protein to load an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo...Development and applications of CRISPR-Cas9 for genome engineering." Cell 157.6 (2014): 1262-1278. PubMed PMID: ...