We narrowed to 804 results for: Eng
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TypeBlog Post...alterations, bringing about a revolution in genome engineering, with new techniques rapidly being developed....and can be used for most of the current genome engineering applications of CRISPR including: generating ... Addgene. Andrew's interests include genome engineering and microbiology. References 1. Chen, Sean...
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CRISPR 101: Making a Knock-In Cell Line
TypeBlog Post...find out! Homology-directed repair and genome engineering The easiest way to make a knock-in cell line ...the right and/or left of the knock-in site. The length of that sequence and the makeup (single vs. double.... A. Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery.... -
Droplet Digital PCR for AAV Quantitation
TypeBlog Post...generating the standard curve for qPCR can be challenging to get right. Here at Addgene, we use a linearized...24328707. PubMed Central PMCID: PMC3991984. Song, Neng, et al. "Detection of circulating Mycobacterium ...droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis." Emerging microbes ... -
Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!
TypeBlog Post...best plasmid for your research can be a daunting challenge. With over 80,000 deposited plasmids from around...That’s why we’ve built a new and improved search engine to help make this task a faster, easier, and frustration-free...sure you’ll find Addgene’s newly launched search engine results much better. Here, I wanted to find a specific... -
Plasmids 101: Protein tags
TypeBlog Post...purify proteins can face any number of technical challenges depending on their protein of interest. After...After several decades of trying to address these challenges, researchers have amassed a considerable molecular...tripled increase the tag size quickly – modifying the length a polyhistidine tract does not greatly alter the... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog Post...pyogenes (SpCas9), is truly an exceptional genome engineering tool. It is easy to use, functional in most species...bacteria) Cas13a targeting with a single crRNA The Feng Zhang lab used Cas13a from Leptotrichia shahii (...cleavage with spacers from 20-28 nucleotides in length and does not require a specific PFS, making it ... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog Post...of 340 institutions with postdocs in science, engineering and health from the NSF’s 2014 Graduate Student...what they are doing, however, may face greater challenges. Current salary information that we have been...current postdoctoral appointments may shorten in length; that postdoc salaries will have to double at some... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post...ICE on Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing field by ...tool is part of Synthego’s full stack Genome Engineering Solutions, which also includes our CRISPR knockout...Browse our CRISPR featured topic page xCas9: Engineering a CRISPR Variant with Pam Flexibility RNA Editing... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog Post...blog post). Optimizing FLP FRT through smart engineering Different FLP versions – making FLP efficient...prevented. The loxP and FRT target sites have been engineered to avoid this re-inversion issue. It became possible...of the asymmetric target site core but its 8 bp length that is critical for Cre and FLP function. Because... -
Hot Plasmids - August 2020
TypeBlog Post...infection (CRISPR-READI) enables genome engineering using HDR donors lengths up to 4.9 kb. Find the plasmids...indicator jGCaMP7, that are optimized for excitation wavelengths above 1,000 nm and enable two-color imaging with... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...PHP.eB were engineered, check out these two references: Chan, Ken Y., et al. "Engineered AAVs for efficient...due to how the Deverman and the Gradinaru labs engineered PHP.B. PHP.B was created by first generating ... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog Post...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...not just wait for papers to be published, but to engage in ongoing, real-time dissemination of information... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog Post...genetic knockout Knock-out cells or animals are engineered by introducing Cas9-mediated frameshift mutations...for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase polymerizes...Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science (80-. ). 368, 290... -
An Integrin Antibody Toolkit from IPI
TypeBlog Post...and significance, these receptors have proved challenging to study. Cells often express multiple integrins...affinity. This extended-open state is key to strengthening ligand binding, mediating cell adhesion and ...adhesion. The results surprised the investigators, challenging the long-held belief that αVβ3 would be the most... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog Post... This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK or in a fast,...Highlights: To detect non-PAM regions, PAM can be engineered into the primers. This method does not depend.../doi.org/10.1101/2020.04.07.028167 Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, Miao... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog Post... the entire length of the cell, even axons of up to 100 microns in length. Some challenges of the Brainbow... -
CRISPR 101: Off-Target Effects
TypeBlog Post...introduces a ssDNA nick, a dsDNA break, or is engineered not to cleave DNA and instead activates/represses...acknowledged as a potential limitation in a study. Strength in numbers If you assess your off-target frequency...Off-target effects in CRISPR/Cas9 gene editing. Front Bioeng Biotechnol. 2023 Mar 9;11:1143157. doi: 10.3389... -
Plasmids 101: Dimers and Multimers
TypeBlog Post...12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on as a dimer, depicted ...tandem, resulting in a plasmid that is twice the length of the monomer (~11.6 kb). Image generated by SnapGene...and fed-batch fermentation processes. Biotechnol Bioeng. 2009 Aug 15;103(6):1129-43. doi: 10.1002/bit.22347... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog Post...technology has made it easier than ever both to engineer specific DNA edits and to perform functional screens...really due to the intended edit rather than some passenger variant that came along with the single cell clone...development of SaCas9, NmeCas9, Cas12a enzymes, and engineered variants thereof offers additional PAM options... -
Important Considerations When Using AAVs
TypeBlog Post...Preps at Addgene 2) Comparison of genetically engineered receptors (i.e. chemogenetics vs. optogenetics...transduction efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities...techniques: exploiting the offspring of genetic engineering." Journal of neuroscience methods 235 (2014):...