We narrowed to 677 results for: Eng
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TypeBlog PostPublishedMarch 25, 2025, 1:15 p.m....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...
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Isolating Genomic Regions of Interest with the CRISPR System
TypeBlog PostPublishedMay 30, 2019, 12:44 p.m....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... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to engineer two new...3) they have limited toxicity. However, a key challenge of using AAVs for neuroscience research is the...AAV.PHP.S targets the PNS since it was actually engineered to target the CNS. How well do these AAVs transduce...takeaways: Two new AAV capsids (figure 1) were engineered with the CREATE method (figure 2) to allow for...References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered AAVs for efficient noninvasive gene delivery ... -
MXS Chaining
TypeBlog PostPublishedFeb. 7, 2017, 3:30 p.m....biological engineering. 2010;4(1):1. Pubmed PMID: 20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia...Knight TF. Engineering BioBrick vectors from BioBrick parts. Journal of biological engineering. 2008;2:5...neighboring fragments require compatible cohesive ends Engler et al., 2008 Gibson Cloning Gibson assembly method...MXS-chaining method have used the technique to engineer various constructs with applications in flow cytometry... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog PostPublishedMay 15, 2014, 3:33 p.m....Aequorea victoria. It has a fluorescent emission wavelength in the green portion of the visible spectrum ...study of dynamic, physiological processes. Re-engineering GFP to increase its range of color and application...its sequence was elucidated, scientists began engineering new versions of GFP through mutagenesis in order...iterations of fluorophores are constantly being engineered. Table 1 lists a few common fluorescent proteins...component (cyan derivatives) Y66H Blue-shifts the wavelength (blue derivatives) Y145F Increases quantum ...at high concentrations T203Y Red-shifts the wavelength (yellow derivatives) A206K Interferes with ... -
Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design
TypeBlog PostPublishedSept. 13, 2018, 1:55 p.m....-targeting cutting as full-length gRNAs Lastly, the effects of guide length on off-target cuts was explored...17-18 bp) have less off-target cutting than full-length guides (19-20bp)? Small-scale studies (Tsai et...explored using full-length (19-20 bp) or short (17-18 bp) guides. When comparing the toxicity of guides...short guides had reduced toxicity compared to full-length guides. Reduced toxicity of short guides could ...28474669. Pubmed Central PMCID: PMC5424143. 2. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide profiling... -
QC Sequencing Technologies at Addgene
TypeBlog PostPublishedJuly 15, 2025, 1:15 p.m.... pairs, long-read sequencing can read sequence lengths in the thousands or even millions of base pairs...BioRender.com. Nanopore can provide longer read lengths of 10 kb to 100 kb (or 4 Mb in ultra-long read ... up the pace Our QC team constantly faces the challenge of keeping up with the large amount of QC analysis...auto-align process to match sequence results to full-length predicted sequences submitted by depositors. A ...100% nucleotide match to base identity and plasmid length. Any mismatch or indel flags the sequence for manual... sequence for a 100% match to base identity and length, but disregards the specified discrepancies. Any... -
Twenty Years of Sharing: Addgene's Viral Vector Service
TypeBlog PostPublishedJuly 30, 2024, 1:15 p.m....one of those stories. Viral vectors are viruses engineered to be safely used to deliver a genetic payload...depending on their serotype, are particularly challenging. Their efficacy in any given system can be affected...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific...activated, respectively, by substrates, light, or engineered ligands (Figure 2). Biosensors, which include...accessible. The supporting data and community engagement required to do this drove Addgene to develop ...partnerships, and new platforms, from our early engagement with UPenn to our launch of the redesigned Data... -
Hot Plasmids - November 2023
TypeBlog PostPublishedNov. 14, 2023, 3:35 a.m....phage-assisted continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, ...(2023). Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell,...delivery and reduce off-target effects of gene therapy, Feng Zhang’s lab has developed a system called DIRECTED...local proteome, but some applications have been challenging or inaccessible. The Ting lab recently made two... increase the utility of this technique. By engineering a light-sensitive LOV domain into the TurboID...organisms. Lee, S.Y., Cheah, J.S., et al. (2023). Engineered allostery in light-regulated LOV-Turbo enables... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog PostPublishedFeb. 17, 2022, 2 p.m....alongside some creative (and significant!) protein engineering. They combined Prime editing with an integrase...right integrase for this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns...potential applications. But they weren’t done engineering quite yet – surely, they thought, the integration...optimized the specific attachment sites via sequence engineering, enabling even more efficient integration activity...system that could integrate genes up to 36 kb in length, with an integration rate of 10-55% - the PASTE...: Adding Precision and Flexibility to CRISPR Engineering CRISPR 101: RNA Editing With Cas13 CRISPR Cheat... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m....Nguyen NT, Zheng Z, Gonzales APW, Li Z, Peterson RT, Yeh J-RJ, Aryee MJ, Joung JK (2015) Engineered CRISPR-Cas9...-associated virus (AAV). At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9) is even...first non-Cas9 CRISPR protein adapted for genome engineering was Cas12a (formerly Cpf1), a nuclease that generates...Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science 368:290–296 . https... genome engineering... -
CRISPR Activation: A Practical Guide
TypeBlog PostPublishedAug. 18, 2016, 2:30 p.m....the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and...that can reassemble when exposed to specific UV wavelengths or chemicals (Polstein and Gersbach, 2015; Zetsche...activator by simply providing a new gRNA rather than engineering an entirely new protein. This makes it much cheaper...the Wyss Institute for Biologically Inspired Engineering. Alejandro (Alex) Chavez is a member of the...Church GM (2015) Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog PostPublishedJune 3, 2021, 1:15 p.m....unfamiliar, scFvs are often the building blocks for engineering proteins. So let’s get acquainted with scFvs....plasmid in bacteria (Ahmed et al., 2012). One challenge of bacterial expression is the proper formation... antibodies to be used as building blocks for engineered proteins. This flexibility is largely due to ...When these molecules, called Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific...minimal systemic side effects. scFvs are part of engineered chimeric antigen receptors (CARs). CARs are a...Bruccoleri RE, Haber E, Crea R (1988) Protein engineering of antibody binding sites: recovery of specific... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog PostPublishedSept. 1, 2020, 1:15 p.m....mediated transduction and expression of the full-length transgene in vitro and in vivo (Allocca et al.,..., the fragments reassemble and produce the full-length gene. Below are overviews and drawbacks of four...and B of the transgene to reconstitute the full-length transgene (Chamberlain et al., 2016). It’s generally...halves of the transgene will reconstitute the full-length transgene. Drawbacks: Trans-splicing occurs...sequence, the 3’ half of the transgene. The full length gene is reconstituted by either the overlapping...10.1038/mt.2013.184 Lai Y, Yue Y, Liu M, Ghosh A, Engelhardt JF, Chamberlain JS, Duan D (2005) Efficient in... -
10 Steps to a Perfect Science Talk
TypeBlog PostPublishedAug. 23, 2016, 2:30 p.m....the best ways to make you and your presentations engaging and memorable. If you read my story above about... results, striking images, creative ideas, and engaging stories as you encounter or experience them. Drop...everything I allow 1 slide for every 1 minute of talk length, and then I cut a few more slides. Do not run over...to be conversational - the audience should feel engaged and not like they’re sitting through a pre-recorded...experience you can experiment with other ways to engage the audience. Take an informal poll by asking for...from the speaker, but actually makes for a more engaging session. The best speakers use no notes and can... -
Evolution of Lab Techniques
TypeBlog PostPublishedJune 21, 2016, 2:30 p.m....all cases, the result can be quantified by the strength of the signal in comparison to a control sample...in the synthesis of DNA strands of all possible lengths because extension is terminated upon the incorporation...become obsolete. For instance, the popular genome-engineering tool, CRISPR, may one day make techniques used...new prenatal test for sickle-cell anemia." New England Journal of Medicine 307.1 (1982): 30-32. PubMed...Andrew Cho, and Ashok B. Kulkarni. "Overview: engineering transgenic constructs and mice." Current Protocols...254.5028 (1991): 59-67. PubMed PMID: 1925562. 10. Feng, Yanxiao, et al. "Nanopore-based fourth-generation... -
Treating Muscular Dystrophy with CRISPR Gene Editing
TypeBlog PostPublishedJan. 26, 2016, 3:30 p.m....Precision genome editing faces the challenges above and more. One chief challenge is upregulating HDR in mature...complex oligonucleotide treatment comes with many challenges, researchers have begun to explore genome editing...necrosis compared to unedited animals. In grip strength, specific force, and other muscle tests, muscle... the time to clinical approval. Even with the challenges precision editing faces, it’s encouraging to ...Central PMCID: PMC3922311. 6. Tabebordbar M, Zhu K, Cheng JK, Chew WL, Widrick JJ, Yan WX, Maesner C, Wu EY... -
Podcast: Interview with Addgene's Founders
TypeBlog PostPublishedApril 13, 2018, 12:51 p.m....Chief Software Engineer at Ginkgo Bioworks. He also holds a SB and a Masters in Engineering from MIT, both...received his Bachelor of Science degree in Computer Engineering from Tufts University and his Master of Arts ...Computer Science. 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... -
Recombinase-based State Machines Enable Order-dependent Logic in vivo
TypeBlog PostPublishedJuly 28, 2016, 2:30 p.m....straight-forward, concise explanation of how to adapt our engineered devices, recombinase-based state machines for...of inputs, may be used to better understand and engineer cellular processes that are influenced by temporally...primer binding sites (designed to have 20-30 nt length, ~60 oC annealing temperature, and ~50% GC content...over a time series of increasing input exposure lengths and found that two hour inductions were sufficient...al. "Serine recombinases as tools for genome engineering." Methods 53.4 (2011): 372-379. PubMed PMID: ...Check Out More Lu Lab Plasmids Find More Genome Engineering Tools ... -
Science Communication Snapshot: DayCon 2015
TypeBlog PostPublishedJune 17, 2015, 9:04 p.m....students develop their communication skills and engage with the public. The organization runs a wide variety...picked a theme (Bio-Medley) to showcase SITN’s strength in the biomedical sciences and decided to offer...offer a varied format event to keep the audience engaged. The day kicked off with introductory lectures ...presenters and board members enabled attendees to engage in small group discussions. The afternoon program...Addgene would love to hear about your efforts to engage the public - let us know in the comments section...