...the recent Keystone Meeting "Precision Genome Engineering and Synthetic Biology". His reflections on the...example, the authors Patrick Hsu, Eric Lander, and Feng Zhang acknowledge the role of data and materials...online user forums such as http://www.genome-engineering.org and http://www.egenome.org." At the Keystone...

...joachimgoedhart.   Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.      Additional... genome engineering...

...becoming an ever more ubiquitous tool for genome engineering, and studying its structure can help us understand... it’s the reason guide sequences have a maximum length of 20 bases: nothing longer will fit. Stabilization...structural insight also spurs the next generation of engineered CRISPR enzymes with their own tradeoffs. And ...the Addgene blog Components of CRISPR  xCas9: Engineering a CRISPR Variant with PAM Flexibility Cas9 vs...

...🎄🎁🎅🤶🎄 pic.twitter.com/rDaEjxTNZX— Heng Chooi (@YitHengChooi) December 4, 2021   I spent a year hoarding...used tip boxes for a lab supply tree and set a challenge to my lab mates. They did not disappoint. #deckthelab...

...completing her PhD in Chemistry and Chemical Engineering with Harry Gray at Caltech, Crystal moved on ... @ShihCrystal.     Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.      Additional...

...synthetic biology, DNA assembly methods, and engineering cellulose degradation. 18:21 - 23:26 : Kwabena's...projects in Ghana.     Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...

...to face global challenges, we hope to empower those that need to face these challenges locally. We look...themselves on the front lines of emerging global challenges. However, these same researchers face significant...

....     Improved nucleic acid sequence detection Feng Zhang’s lab recently developed a platform called...efficiency and specificity of multiple wild-type and engineered variants of Cas nucleases. In doing so, they ...created a modular toolkit for Cas-mediated genome engineering in plants. Read the paper in bioRxiv Find...

... the Collins' lab at the time) got the idea to engineer bacteriophage (viruses that infect bacteria) for...grow with his help! Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...

... up with it. The peer review process has been challenged with many closely spaced CRISPR software submissions...facilitate most experimental applications in CRISPR engineering. When compared to that collective utility, it...This is only one of many decisions faced by those engaging in CRISPR design, but it is a significant hurdle...limitation, but the tool is fast enough that, if you’re engaged in heavy CRISPR design (I’m talking screens and...

...My son is an engineer and my daughter has a business degree. From the very start of their undergraduate...talent in science related careers. Failure to strengthen the talent pipeline sets the world up for reduced...Image from pd|hub.   To begin to tackle these challenges, a group of stakeholders came together with the...

...exciting developments surrounding CRISPR genome engineering in the past year, we're excited to announce that...lentiCRISPR V2   LentiCRISPR v2 originates from Feng Zhang’s lab at the Broad Institute and was designed...PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide libraries...

... some challenges during my internship, but nothing could have prepared me that those challenges would ...exploring my different interests but also a way of challenging what I am used to. Brandon Solis, marketing...

...genome edits compared to rat APOBEC1. Now, they’ve engineered SECURE adenine base editor variants using a protein-truncation...activation-induced cytidine deaminase (AID) and an engineered tRNA adenosine deaminase (TadA) to a catalytically...packaging plasmids   Cytosine deaminase for RNA editing Feng Zhang’s lab previously developed a programmable ...

...be removed by re-expressing the transposase. A challenge of using PB to generate iPSCs is that genomic ...the cytoplasm independent of the cell cycle. A challenge of using Sendai is that since it’s replication...clones tested had plasmid integration. Another challenge is that multiple transfection makes it difficult...induced pluripotency: Recent progress and future challenges." Journal of Cellular Physiology 228, no. 2 (...

...co-op) is what has brought me, a second year bioengineering student, to Addgene. I am currently working...interview. I anxiously prepared for the call. The length of this interview was intimidating to me, as I ...comfortable.     Lucie Wilson is a rising junior and bioengineering student at Northwestern University. She is ...

...researchers involved. Communication is a major challenge when researchers are communicating across disciplines...disciplines or doing different tasks. But these challenges can be exaggerated when communicating across ... help other teams successfully overcome these challenges, too. Schedule regular meetings Scheduling a...

...them attractive tools for the life scientist or bioengineer.   Figure 1: Map of a plasmid with its...reporter plasmids, viral plasmids, and genome engineering plasmids. Some of the many things that plasmids...controlled changes to an organism’s genome (genome engineering) Produce synthetic viruses that can be used in...

.... The resultant JEDI-2P sensors overcome many challenges in 2-photon in-vivo imaging of voltage fluctuations...neurons, far out-performing previous attempts to engineer potassium-selective channelrhodopsins. HcKCR1 ...work also proposes promising avenues for future engineering to shift the spectra and improve the performance...

...the Rluc8 was more challenging. The Roth lab used structure-guided protein engineering and experimental ...the blog post. References Olsen RHJ, DiBerto JF, English JG, Glaudin AM, Krumm BE, Slocum ST, Che T, Gavin...

...fluorescent protein-based sensor. This sensor was engineered from a GFP variant and a DA receptor such that...PMCID: PMC3083031. Kebabian, John W., and Paul Greengard. "Dopamine-sensitive adenyl cyclase: possible ... Kebabian, John W., Gary L. Petzold, and Paul Greengard. "Dopamine-sensitive adenylate cyclase in caudate...

...cycle of one step. Challenges in the LEGO science video process One major challenge when we started was...will bring people together and make these daily challenges a little easier to overcome. We want to encourage...

...Kurt Swanson a structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in...FOP until a cure is available. Addgene and AJ's Avengers This year Addgene, where my wife Lianna has made... Swanson is a structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in...

... fiber (Park et al., 2019). This provided an engineering road map of sorts, giving an idea of the stiffness...whenever necessary. This is all an impressive engineering feat, but you’re probably wondering “did it actually... Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC), 4322–4325...

...Enhancement in vector stability." Biotechnology and bioengineering 94.2 (2006): 322-329. PubMed PMID: 16528756...pressure and sodium butyrate." Biotechnology and bioengineering 42.5 (1993): 601-610. PubMed PMID: 18613082...and antibody production." Biotechnology and bioengineering 37.10 (1991): 989-993. PubMed PMID: 18597326...

...could potentially be used to report upon and/or engineer “temporally ordered biochemical events” such as...RNA-based toehold switches described in this post are engineered to bind RNA sequences derived from Zika virus..., Multitasking “CRISPR-OS” In this post, Albert Cheng and Mark Wanner from the Jackson Laboratory for ...

...remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C. elegans were released, signaling...revolution!   References Streamlined Genome Engineering with a Self-Excising Drug Selection Cassette....strategies for CRISPR/Cas9 editing in C. elegans: Engineering the Caenorhabditis elegans genome using Cas9-...

...since the very beginnings of the field. Those engineered sensors were designed to replace synthetic calcium...problem that he considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors...trial and error. Transition metals also present challenges in that they are found in cells at much lower...

...exploration site Inventing Heron has a radio reporter challenge, and code.org has coding tutorials for all ages...environment.  Be kind to yourself. This is a challenging period for everyone for numerous reasons, and...check out She Geeks Out, Science, or Resources to Engage Kids at Home. It’s normal and okay to feel overwhelmed...

...sources of information if those sources affirm or challenge their beliefs. I asked participants to read short...read an abstract for a study with findings that challenged their beliefs, they were much more likely to ...dissonance (Kunda, 1990). More specifically, they were engaging in a behaviour called the “biased evaluation of...

...  7 ways to strengthen your new relationships Here are some ways to go about strengthening a new relationship...often the easiest way to build relationships and strengthen them over time.  Things to avoid when following...

...initially limit the technology to research centers engaged in long-term, mandatory follow-ups and observational...Download the CRISPR 101 eBook Read our genome engineering blog posts Resources on Addgene.org Read our...CRISPR plasmids by function Check out our genome engineering guide ...

...recombination-mediated genetic engineering (find more information on bacterial engineering using CRISPR on our blog...interactions. Combining synthetic biology with engineering: glowing bacteria and a fish robot to detect ...

...newborn gut. One of Fatima’s ultimate goals is to engineer bacteria for the efficient product of HMOs that...surfaces. After incubating the sensor with bacteria engineered to produce HMOs, sensor binding results in an...future work with HMOs will enable researchers to engineer healthier microbiomes in people all over the world...

...seqWell, Inc., our mission is to overcome crucial challenges in NGS by developing technologies that can help...problem in multiplex NGS library preparation. The challenge here is that, as more samples are sequenced simultaneously...enhanced normalization of read count and fragment length across samples, NGS data obtained via plexWell™...

...Cofactors/Substrates Required Not Required Signal Strength Lower Higher Sensitivity Higher Lower Background..., substrate requirements and photon emission wavelengths. Renilla luciferase from the sea pansy (Renilla...these luciferases have been subjected to further engineering to improve upon the properties of the native ...

...many career pathways exist for scientists and engineers, a few brave scientists will choose to start their... interdisciplinary, with emphasis on genetic engineering, sequencing, and translation of results to companies....” Take home message Starting a lab can be a challenging process. Each PI emphasizes the importance of...

...Non-Continuous Evolution (PANCE) selection systems to engineer better and faster ABEs (Fig. 2). These systems...ABE7.10. Find the ABE8e plasmids at Addgene! Why engineer and optimize base editors? Targeted evolution ...Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR (2020) Phage-assisted...

...fluorescent as well as FRET-based biosensors have been engineered to detect a wide array of neurotransmitters ranging...biosensors? Well, a lot of these sensors are already engineered into viral vectors, so if you want to use them...Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in Cellular Neuroscience...

...and tractable tools for genome engineering. Scientists have engineered viruses to target specific cell...infect. In the lab, this means viruses can be engineered to be cell type specific. Once inside the host...

...reacting to a chemical, but by emitting a specific wavelength of light in response to excitation by a laser... degrading them. You’ll also need to know the wavelength of laser needed to excite them - and it’s a good...   Fig 2: Try to inversely pair fluorophore strength with protein abundance.    Second, you’ll ...

...pGGA008 (AlcA) and pGGC011 (AlcR) from red flame GreenGate kit Steroid-regulated LexA promoter XVE (synthetic...shock-inducible Cre and Cas9 for easy genome engineering in species like C. elegans and Drosophila. Light... induce transcription of your target gene? The strength of inducible promoters can vary a lot. If you’...

...Immunologists are coming together to tackle the challenge of curing cancer in cross-disciplinary projects...CRISPR/Cas9 methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor..., CRISPR/Cas9 genome editing allows targeted engineering of the TCR locus. He also highlighted the UCSF...

... on differences in emission wavelength. Filtering the emission wavelength partially separates the signals...In fluorescent imaging, you use the excitation wavelength to specify which FP emission you’ll capture. ...

...this virus. While various Cas9 mutants have been engineered to perform unique editing tasks, classic Cas9...LentiCas9-blast is a Cas9 expression vector developed by Feng Zhang’s lab at the Broad Institute (Sanjana et al...makes generation of high titer Cas9 lentiviruses challenging. This cytotoxicity also causes growth rate variability...

...computation work comes with new sets of challenges. One challenge with computational analyses is to keep... are doing what you are doing. Another common challenge is to keep track of what works, and what does ...

...advantage to using the CRISPR/Cas system for genome engineering is the fact that Cas9 can be easily programmed...smegmatis (Yan et al., 2020). NHEJ can also be engaged by variants of the canonical Cas9 approach. A pair...UNC Chapel Hill.   References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM (2009) Limiting the Persistence...

... is a postdoc at Harvard University where she engineers genetically encoded voltage indicators. Tian developed...mutant sensors expressed in a mammalian cell line engineered to be electrically excitable. The mutants are.... Tian notes that this platform can be used to engineer many types of biosensors.   This screening method...

...injected with an immunogen. This might be the full-length protein that you want to generate antibodies against...Otherwise, you might experience variability in the strength of staining and background noise. If you do use...control across experiments to be aware of signal strength. In general, polyclonal antibodies produce a higher...

... evaluate degradation of ENL. Cell lines were engineered to express ENL-FKBP12F36V and endogenous ENL ...evaluate degradation of MELK. Cell lines were engineered to express FKBP12F36V-MELK and endogenous MELK... evaluate degradation of YY1. Cell lines were engineered to express YY1-FKBP12F36V using CRISPR/Cas9-mediated...