...point: Test different lengths of the primer binding site, starting with a length of about 13 nt (Anzalone...optimized (Anzalone et al., 2019). Test different lengths of reverse transcriptase template, starting with...templates, it is even more important to test different lengths and sequences, as unintended secondary structures...Anzalone et al., 2019). Figure 2: Starting lengths for optimizing your pegRNA primer binding site ...Editing the PAM prevents the prime editor from re-engaging with DNA it has already edited. Created with BioRender.com...., Chen, J. C., Hsu, A., & Liu, D. R. (2022). Engineered pegRNAs improve prime editing efficiency. Nature...

...(7629):452. doi:10.1038/nature20149 Jiang Q, Meng X, Meng L, et al. Small indels induced by CRISPR/Cas9...RNAs (lncRNA). lncRNAs are over 200 nucleotides in length and lack an open reading frame, so small deletions...promoter and enhancer sequences remain intact (Engreitz et al. 2019). The interactions between ncRNAs ...RNA-targeting CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer...18(12):2097. doi:10.1080/15476286.2021.1899500 Engreitz JM, Haines JE, Perez EM, et al. Local regulation...in Plants with the CRISPR-Cas Technology is a Challenge yet Worth Accepting. Front Plant Sci. 2015;6. ...

...elegans Fluorescent timers for studying cell cycle length The CRISPR corner New from the viral service ...synaptic connections and physiology is particularly challenging as they are great in number, generally very small...come together, they fuse and fluoresce. The team engineered C. elegans cell lines to express wrmScarlet1-... small wrmScarlet11 which is less than 60 nt in length to tag the target protein. The team validated this...10.1101/2020.07.02.185249 Predicting cell cycle length with a histone-fused fluorescent timer By Kate ...fusion protein can be used to determine cell cycle length by examining the ratio of blue to red cells. Cells...plasmids at Addgene Sc++ is a codon-optimized, engineered Streptococcus canis Cas9 nuclease that has a ...

... Centre for Public Engagement (NCCPE) provides on-site courses in public engagement for institutions and...science blogs like @WIRED Science. Don’t be scared to engage with them. Comment on their activities and participate...using social media as a scientist here. 2) Public engagement - Participate in science festivals and outreach...talking in public, volunteer at science festivals. Or engage with non-scientists in an unusual setting: Discuss...by advertising your blog on social media and by engaging with other science bloggers. If you don’t have...guest blogger, Sarah Schmidt! Sarah Schmidt is engineering resistance against Fusarium wilt disease in bananas...

...since the Future of Research Symposia Ongoing challenges facing the scientific enterprise and the way ...revealed that young scientists are aware of, and are engaged in, these issues which will continue to shape the...landscape of the scientific enterprise will provide challenges and opportunities for us all.  For example, implementation...categorize all postdocs at institutions, will be challenging. At the same time, there is much opportunity ...the pressure on for further increases will be challenging, but will be aided by the development and implementation...science funding and funding mechanisms require an engaged sector with effective advocacy skills. Advocating... is an important way to push the balance from challenge to opportunity. We hope to see you at the upcoming...

...of Revolution Bioengineering.  Last year we started a company. Revolution Bioengineering is two responsible... We both had an applied science bent, so the engineering aspect of this job appealed to us. However, since... publish or attend conferences makes it very challenging to compete in today’s tough academic or industry...team. We have confidence that we can take on any challenge and quickly become experts in something, even ...comment.  In our day job, we worked on plants engineered to detect explosives and rapidly indicate their...careers: to build something new and beautiful, to engage with the public in a meaningful way about science...

...xkcd.com Science communication should be fun and engaging for the communicator as well as the audience, ...enjoy and that plays at least slightly to your strengths. Otherwise, you’re going to drop your project ...graphics for visuals. However, it’s much more challenging to design a hands-on interactive activity using... Small tidbits and fast/fun facts are easy and engaging on social media, but deep dives are often more...complexity. Podcasts Medium Easy for audience to engage while multitasking. Easy to host guests remotely...etc… Low (except for space) Real-time audience engagement and response. High connection levels and interaction...geographic location) Opportunity for real time engagement while allowing for recording/future access. Remote...

...those engineered DNA-binding molecules for the first time and named the new technology as engineered DNA-binding...insertional chromatin immunoprecipitation (iChIP) and engineered DNA-binding molecule-mediated chromatin immunoprecipitation... locus, which is the rate-limiting step. When engineered DNA-binding molecule technologies such as TAL...However, it struck me that direct use of such engineered DNA-binding molecules even enables us to skip...Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation... and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin immunoprecipitation...

...studying chemical engineering. Mengtong "Tom" Duan is a PhD student studying bioengineering. They are both...agents. It ended up being very straightforward to engineer various cell lines from different tissues and ...transposase. In rtTA-cell lines, it is better to first engineer the cell line with rtTA, then integrate the mARGs...was sufficient to saturate expression. When engineering a cell line, cotransfect the mARG cassettes 2...For in vivo experiments, simply induce injected engineered cells via regular intraperitoneal doxycycline...

...solutions to complex challenges. As a cross-disciplinary researcher, the challenge of keeping up on studies... Sharing your research with the world can be challenging. After months, or years, of grueling effort to...biology or international relations.  Many of the challenges in science today are too complex for any single...looking for funding, or are by researchers for whom English is a second or third language (making publishing...publishing their research all the more challenging even when it is of quality). On the site you can, for example...Vocalization. The video summarizes the research of Drs. Feng and Bass published in a recent issue of the open...

...despite being approximately half the length (Warner et al., 2014); its length reduces artefacts during live ...RNA is not inherently fluorescent and can be challenging to track intracellularly, but aptamers permit...vivo. However, Spinach has proven useful in the engineering of riboswitches. Riboswitches are naturally occurring...Nilaratanakul et al., 2017) and levels of the second messenger cyclic di-AMP in Gram-positive bacteria (Kellenberger...than any form of Spinach - only 49 nucleotides in length - and is better able to maintain its fluorescence...fluorescent biosensors for live cell imaging of second messengers cyclic di-GMP and cyclic AMP-GMP." Journal of...

...water, which absorbs wavelengths above 900 nm or so. In other words, wavelengths between 650 nm and 900...animal and human tissues largely unimpeded. The challenge then was to develop fluorescent proteins that ...spectral properties of PA FPs with light of specific wavelength and intensity allows for optical labeling and... scattered,” Verkhusha explained. Ultrasound wavelengths, on the other hand, are larger than the size ...photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome. Piatkevich et al...

...correlated to the length of the truncated attP site. The three SPARC variants, based on the length of the attP...doi.org/10.1101/788679 Lin R, Wang R, Yuan J, Feng Q, Zhou Y, Zeng S, Ren M, Jiang S, Ni H, Zhou C, Gong H,...titration of the AAV. So far, tools to overcome this challenge in Drosophila require heat-shocking the system...Learn about Gateway cloning Read about genome engineering in flies Learn more about controlling gene expression...

...associated genes) system. This popular genome engineering tool offers flexibility, multiplexibility, and... DNA into small fragments approximately 30bp in length and paste them into the CRISPR sequence. Separate...system that scientists have harnessed for genome engineering. The diversity of the CRISPR/Cas systems provides...defense against invading phages. Rising to the challenge - Evolution of anti-CRISPR genes in phages The...who has the strongest will to survive. Genome engineering and future directions CRISPR/Cas inspired genome...Despite the ubiquity of CRISPR/Cas toolkit, genome engineers continue to work diligently and creatively to ...

...impossible. With the genome engineering revolution came epigenome-engineering tools - zinc finger nucleases...Wells D, Ahmed AA, Sauka-Spengler T (2017). Genome and epigenome engineering CRISPR toolkit for probing...the DNA after cytosine removal. Recently, Albert Cheng’s lab developed Casilio-ME, which is based on their...Lysine-specific Demethylase 1 (LSD1) Tatjana Sauka-Spengler's lab has deposited pX330a dCas9-LSD1 for targeted...2017) Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nature Communications...

... sequence, making it very useful for genetic engineering approaches. Most importantly for this technology...CRISPR/Cas9, TALENs, and ZFNs). However, several challenges still remain, such as making gene edits completely...nuclease technology with the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon...experience with stem cells to Transposagen's cell engineering team.     Mathew Goodwin is a Technical Sales...Craig, “piggyBac transposase tools for genome engineering.,” Proc. Natl. Acad. Sci. U. S. A., vol. 110,...Sleeping Beauty Transposase Read up on Genome Engineering with FLEx Vectors  Learn how Homology Directed... genome engineering...

...neuroscience—whatever that entails. Despite being engrossed by the theory and findings of brain science, my... to once again write for fun eventually saw an English minor tacked onto my BSc. With this decision I ...meaningful day job only came into focus when an English tutor noted my eclectic mix of science and arts...so—can prove invaluable in the pursuit of public engagement. Studying science communication is an equally...had to say. Moreover, this subject allowed me to engage in findings relevant to my background (neuroimaging... and the tools to deploy its wisdom remains a challenge. Departments like ours reflect a growing effort...

...classroom Engineering the Plant Genome Using CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources...Arabidopsis in the classroom has many benefits. Engaging students in active learning through hands-on experiments...system comes with fewer ethical and logistical challenges than does working with animals. Because Arabidopsis...and share resources, we seek out opportunities to engage people in science learning. Whether visiting classrooms...University. In her work, she focuses in public engagement in science and promoting plant science in K-12...

...100% student engagement. I figured, an engaging card game could help me get the engagement I wanted. I ...Associate Professor at University of Barcelona. The challenge: Making courses on genetic manipulation more hands...could be played when and, depending upon who your challenger was, you might have to drastically alter your...biology, cell biology, microbiology, genetic engineering and strategic knowledge to solve a clinical disease...

...Novel of England by Edward Rutherfurd "A 10,000 year history of southern England. While the length may be...fiction. Two dueling magicians in the 1800s in England who help bring back actual magic (which had been...one. On a purely selfish level, I wanted to be challenged to read some books outside of my comfort zone...life. Though I have an undergraduate degree in English, I had lapsed. Baker’s novel reignited my appreciation...

... of the few members of Congress who holds an engineering degree. It was a great, positive event that I...a PhD Candidate studying Nuclear Science and Engineering at Rensselaer Polytechnic Institute. He also ...groundbreaking research happens, sometimes forgetting how to engage with friends, family, and the public about its ...policy.  He is also chair of the postdoctoral engagement committee and active member of the university's...present at the lab bench," Liu, a UM undergraduate, challenged us. I worried the march would be a simple, unquestioning...scientists must rise up and become more politically engaged. I was proud to be a part of a community that recognizes...face of evidence. Stephanie Hays is moving from engineering synthetic microbial communities at Harvard University...

...expression to a specific neuronal subtype. However, the length of promoter that can be used in an AAV construct... As an alternative, some constructs have been engineered with a double-floxed inverse open-reading frame...transgenic mice Transgenic mouse lines have also been engineered to express various opsin genes [7-9]. The advantage... cell body or an axon terminal also affects the length of time you may need to allow the virus to express...opsin to express and passively diffuse down the length of the fiber before stimulation of the terminals...and must passively diffuse so, depending on the length of your projection, it may take as long as 6-8 ...9):745-52. PMID: 21985008; PMCID: PMC3191888. 9. Zeng H, Madisen L. Mouse transgenic approaches in optogenetics...

...for many metabolic engineering and synthetic biology goals. In metabolic engineering, the expression of...plasmids in order to study gene function or to engineer cells for specific purposes. Unfortunately, plasmid...interfere with the precision needed in metabolic engineering or other synthetic biology circuits. What is ...Eduardo D. Sontag, and Christopher A. Voigt. "Engineered promoters enable constant gene expression at ...coli using DIAL strains." Journal of biological engineering 5.1 (2011): 10. PubMed PMID: 21787416. PubMed...

...reprogrammed regenerating tissues without using genome engineering. Planarian worms cut into three segments “remember... wild type  (Oveido et al., 2010). One of the challenges of studying bioelectricity is that it can be ...between open and closed states using different wavelength light pulses. Other resources from the bioelectricity...defects, regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use...growth and form." Annual review of biomedical engineering 19 (2017): 353-387. PubMed PMID: 28633567. Mathews...bioelectricity for regenerative and synthetic bioengineering." Current opinion in biotechnology 52 (2018...

...biological systems can hinder our attempts to study and engineer them, but what if we had a simple tool that allowed...phenotype, and quickly profile the resultant hits (Cheng et al., 2014; Wong et al., 2015; Wong et al., 2016...genetic elements including transcription factors (Cheng et al., 2014) and microRNAs (Wong et al., 2015)....can be contacted at [email protected].   References 1. Cheng, Allen A., Huiming Ding, and Timothy K. Lu. "Enhanced...Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex." Nature (2014). PubMed PMID...CRISPR Plasmids Find More Tools for Combinatorial Engineering on Our SynBio Pages ...

... while engineering viruses Acr genes can be used as positive selectable markers in engineering viruses... was used to replace genes in the difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz...that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-Cas system. 5. ... proteins are small enough that they could be engineered to “armor” therapeutic phages to protect them...He F, Jørgensen J, Madsen P, Bhoobalan-Chitty Y, Peng X (2018) Anti-CRISPR-Based and CRISPR-Based Genome...

... cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery technologies have...adenovirus for productive infection. AAV vectors are engineered to provide additional safety benefits: they lack...treatment appears to be both safe and effective. Challenges for AAV gene therapy Although AAV-based gene ...promising, scientists are working to overcome key challenges to their efficacy:  Pre-existing immunity: Many...with gene therapy in childhood blindness." New England Journal of Medicine 372.20 (2015): 1920-1926. PubMed...PubMed PMID: 12756417. 5. Chan, Ken Y., et al. "Engineered AAVs for efficient noninvasive gene delivery ...

...) signal. While many studies focus on promoter strength as a determinant of gene expression levels, the...multiple adenine (A) nucleotides to the tail of a messenger RNA transcript. The purpose and mechanism of polyadenylation... terminators, such as T7 and rrnB, as well as engineered high-efficiency terminators such as T0. Rho-independent...annotation for 90 bacterial elements. By Ppgardne at English Wikipedia, CC BY-SA 3.0, https://commons.wikimedia.org...initiating the desired cleavage event, although some engineered terminators come close (>95%). For most purposes...detailed analysis of yeast terminators of varying strength. A discussion of the effects of various terminators...

...advs.202202957 Yuan, T., Yan, N., Fei, T., Zheng, J., Meng, J., Li, N., Liu, J., Zhang, H., Xie, L., Ying...researchers in the Keith Joung lab, who were engineering an ABE to reduce its tendency to make off-target...they were using consisted of Cas9 fused to an engineered adenosine deaminase capable of chemically converting...sites. To explore this further, the team started engineering a CBE to be worse at its original job. They ...causing double-strand breaks (DSBs). The Fei-Long Meng Lab did in-depth profiling to compare editing outcomes..., Wang, B., Luo, Y., Cai, Y., Liu, X., Xu, Z., … Meng, F.-L. (2024). C-to-G editing generates double-strand...

...Fujita and Fujii, 2013). They call this method engineered DNA-binding molecule-mediated chromatin immunoprecipitation...immunoprecipitation (enChIP) because any engineered DNA-binding molecule, such as Zinc-finger or TAL proteins...the "in cell" form of this method, in which an engineered DNA-binding molecule such as the CRISPR complex... and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin immunoprecipitation...Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation...genomic regions." Journal of bioscience and bioengineering 108.5 (2009): 446-449. PubMed PMID: 19804873...