...enzymes are the latest tools in the transcriptome engineering toolbox. How were Cas13d enzymes discovered?...property is size - Cas13d enzymes have a median length of ~930aa - making them 17-26% smaller than other... tell how large the genome and transcriptome engineering toolbox will be. It is, however, certain that...References Konermann, Silvana, et al. “Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors...

...method that can be used for cloning or genome engineering and is based on homologous recombination. It ...lambda red bacteriophage and its use as a genetic engineering tool is frequently called recombineering - short...for homologous recombination-mediated genetic engineering.  It can be used to make an assortment of modifications...efficient recombination system for chromosome engineering in Escherichia coli. PNAS 2000 97 (11) 5978-5983...Homologous Recombination-Based Method of Genetic Engineering. Nature protocols. 2009;4(2):206-223. doi:10.1038...and Court, D. L. 2014. Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination. Current...Mammalian Cells CRISPR Methods for Bacterial Genome Engineering Resources on Addgene.org Browse our Microbiology...

...by Benoit Giquel. CRISPR has taken the genome engineering world by storm owing to its ease of use and utility...Waltz, 2016), the potential of CRISPR for genome engineering in plants is also being realized. There are a...plants with the phenotype of interest. Genome engineering, on the other hand, allows for targeted modification...phenotype. In fact, CRISPR has already been used to engineer the genome of many plant species, including commonly...will present a general overview of plant genome engineering using CRISPR, highlight the specific modifications...plants, and outline the various plant genome engineering tools that are available to academic researchers... Addgene. CRISPR components for plant genome engineering CRISPR can be used to knockout, activate or repress...

...students (1, 2), but educators face significant challenges in designing suitable projects. In the O'Connor...Advisors on Science and Technology [PCAST] (2012). Engage to Excel: Producing One Million Additional College...Graduates with Degrees in Science, Technology, Engineering and Mathematics, Washington, DC: US Government...

...career path and why he chose to go into genome engineering. Chapter 1: Early research on CRISPR and what... 6: Projects in Wei Leong Chew's lab. Genome engineering, gene delivery methods, AAVs, and more. Chapter...and gene therapy.   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...

...bacterial CRISPR engineering One common way of accomplishing bacterial genome engineering is done with recombineering...the genus Streptomyces, yet genome engineering remains challenging for most organisms in the genus. Work...discovered in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria...quirks change the way CRISPR-mediated genome engineering functions in bacteria, but have no fear - plasmids...recombineering-based MAGE (Multiplex Automated Genome Engineering) technology (Ronda et al., 2016). pMA7CR_2.0 ...E. coli and T. citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines...create a system for scarless, iterative genome engineering (Jiang et al., 2015). pCas contains Cas9 and ...

...system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan...making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that gap. As Sakuma and...to include even more loci. Multiplex genome engineering demonstrated in human cells They used the Golden...efficient viral vector-mediated multiplex genome engineering. The system now provides an efficient targeting...Sakuma’s protocol. Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector... multiplex genome engineering...

...post-transcriptional regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed...has opened up a world of possibilities in the engineering of complex gene networks. A comparable system...assemble designer PUF domains for RNA specificity engineering. “The RNA binding domain is interesting because...report published in The Journal of Biological Engineering in March is fused to a translational repressor...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...molecular definition, and potential for further engineering as compared to polyclonal or monoclonal antibodies...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...

...within tumours via focused ultrasound. Nat Biomed Eng. 2021 Nov;5(11):1336-1347. doi: 10.1038/s41551-021...antibody?  Similarities:  Generated against full length GFP and converted to recombinant form by the NeuroMab...these processes. The Liu and Petsko labs recently engineered hyperfolder YFP (hfYFP), a FP well-suited for...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...empower the scientific community with the tools to engineer them. Given the ease with which scientists can... Gong B, Nguyen K, van der List D, Murray KD, Engebrecht J, Trimmer JS (2019) A toolbox of IgG subclass-switched...Monoclonal Antibodies for Clinical Use. In: Antibody Engineering. InTech Additional resources on the Addgene blog...

...their applications in neuroscience. Curr Opin Biomed Eng. 2019. 12:111-117. doi: 10.1016/j.cobme.2019.10.010... as bright as the other GEVI systems that use engineered FPs. Attempts have been made to optimize their...their fluorescence, which ultimately led to the engineering of the second type of these GEVIs: a FP-fused... voltage sensor. FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component... transiently and for genomic integration. Lab-engineered viruses will package your genetic cargo in lieu...

...pluripotency and nuclear reprogramming. Adv Biochem Eng Biotechnol, 2009. 114: p. 185-99. PubMed PMID: 19343304...embryonic stem-cell lines from human blastocysts. N Engl J Med, 2004. 350(13): p. 1353-6. PubMed PMID: 14999088...

...of mice via adeno-associated viruses. Nat Biomed Eng 4:97–110 . https://doi.org/10.1038/s41551-019-0501...gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb... little room for additional regulatory elements. Feng Zhang's group previously packaged Cas9 and multiple...enough to package into AAV. At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9) is the...ADAR2DD(delta984-1090) are approximately 4.1 kb in length, allowing them to be packaged in AAV.   Split ...transgenes might seem daunting, it’s actually a challenge that the field has overcome with split AAVs. In...When a cell is transduced by both AAVs, the full length gene and/or protein is reconstituted. One way to...

...enabling technologies in synthetic biology. J. Biol. Eng. 2013; 7, 13. PubMed PMID: 23663447. PubMed Central...fragments of the chromosome, which was technically challenging. Other homology based technologies We've described...Central PMCID: PMC3874208 Other Resources: New England Biolabs Gibson Assembly Overview Additional Resources...Protocols Gibson Assembly® is licensed to New England Biolabs by Synthetic Genomics, Inc. ...

...mechanobiology: seeing is believing." Annu. Rev. Biomed. Eng. 10 (2008): 1-38. PubMed PMID:18647110. Additional...spectral properties upon exposure to a specific wavelength of light. PA-FPs can be activated from low fluorescent...brighter fluorescent state by exposure to a specific wavelength of light. The switch or photoconversion usually...multiple times by activating with two distinct wavelengths of light (see Table 2). The best known reversible...photoactivation) (6). Dronpa is photoactivated by blue wavelengths of light, causing emission of green fluorescence...another group of reversible PA-FPs. These were engineered to be the first reversible, monomeric red fluorescent...dynamics (fission, fusion, etc). For example, by engineering PA-GFP tagged to a mitochondrial matrix protein...

...2014. PubMed PMID: 24151867 Mikkelson, et al. Metab Eng. 2012. PubMed PMID: 2232647   SapTrap, a toolkit...genome engineering across multiple species Recent discoveries in the fields of genome engineering and synthetic...pCAGGS-TAL-NC2 Charles Gersbach pcDNA3.1-GoldenGate, pcDNA3.1-GoldenGate-VP64 Maria-Elena Torres-Padilla pTALYM3...fold a protein because of its length. Mark Howarth’s lab recently engineered a new peptide/protein pair,...as not only a protein engineering tool, but also as a way to characterize/engineer promoters, enhancers... developed for the study of enhancer function, strength, and binding (e.g. DHS-seq, FAIRE-seq, and CHIP-seq...quantification of enhancer activity have remained challenging due to the variable availability of enhancer ...

...ways of delivering healthcare. Clients typically engage consultants as short-term experts to work on “cases...Cases present specific operational or commercial challenges or questions that clients face imminently. Consulting...on analyzing the situation that presented the challenge at hand and help resolve it through well-informed...specialize in business strategy tend to be mostly engaged by pharma and biotech clients. In this realm, there... there are a few different kinds of challenges that consultants get called upon to tackle, depending on...to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...rewarding parts of the job is being confronted with challenges that are crucial for the success of a drug’s ...

...accomplish is to make the engineering of biology much easier. So you could imagine engineering the entirety of ...dig into its engineering qualities, what these parts allow is for someone to forward engineer a system of...Drew Endy's lab at Stanford develops engineered DNA systems capable of storing data and computing inside...an organization working to advance biological engineering openly so as to benefit all people and the planet... to not assert property rights for particular engineered genetic sequences; but it’s only the promises... what we think of as an operating system for engineering an entire cell. Typically when people use plasmids...of a cell. That's a big long-term technical challenge. We have a strategy that organizes how we start the...