We narrowed to 114 results for: N-pac
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TypeBlog PostPublishedJuly 15, 2014, 4:03 p.m....regulation. Molecular Cell 27, 691-700. (PubMed) McKenna, N. J.and O'Malley, B. W. (2002). Combinatorial control...cascades conceivably allows the genome to sense the impact of the total environment on the cell. The NURSA...
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Overcoming the Challenges of Lentiviral Production
TypeBlog PostPublishedAug. 7, 2018, 12:31 p.m....Kumar M, Keller B, Makalou N, Sutton RE. Systematic determination of the packaging limit of lentiviral vectors...generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes VSV-G, a packaging plasmid...lentiviral packaging systems please see Addgene’s Lentiviral Guide. While the third generation packaging systems...requirement for additional DNA can impact the transfection efficiency of the packaging cell line and thereby affect...are transfected into a packaging cell line, usually a derivative of 293T. Packaging cells are incubated ...delivering genes via lentivirus is deciding what packaging system is best for a particular experiment. The...The newer generation lentiviral packaging systems were designed to prevent the formation of replication... -
Antibodies 101: Affinity Reagents
TypeBlog PostPublishedApril 14, 2022, 1:15 p.m....Chemistry. https://doi.org/10.1074/jbc.M407888200 Kwon, N., Kim, Y., Lee, J. (2019). Structural diversity and...amazing affinity reagents we just didn't have the space to give proper credit to. Thankfully, as Addgene's..., or be able to access an epitope folded into a space that would size-exclude an antibody. In imaging ... -
Antibodies 101: Fab Fragments
TypeBlog PostPublishedSept. 5, 2024, 1:15 p.m....Sakai M, Tomisaki M, Kiuchi T, Takagi J, Watanabe N (2022) Engineered fast-dissociating antibody fragments...sometimes you want a tool that is a little more compact — more of an antibody fragment rather than the ... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O, Chambon P, and Ghyselinck ...and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific Recombinases on Genetic Analyses... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog PostPublishedDec. 10, 2020, 1 p.m....Vidigal JA, Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo...provide a large transgene packaging capacity. For instance, with a packaging capacity of about 6 kb, the 1st...of AdVs have already a packaging capacity larger than AAV vectors. This capacity is large enough to carry... -
A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog PostPublishedMay 7, 2020, 1:15 p.m....to contain a specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can be any nucleotide... Depending on the position in the protospacer, the editing capacity of the ABE8s ranged from 1.5x higher...ABE8s demonstrate an overall improved base editing capacity, even at sites that used to be difficult to target.... References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable...Edwards A, Gehrke JM, Lee S-J, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog PostPublishedJune 12, 2014, 3:30 p.m....plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is a tobacco relative and a good model...other people to do that too. We don’t have the capacity to send tools out to people ourselves, but we’...committees, and how the culture of the university impacts the number of women reaching senior positions. ... those amazing students that want to have a big impact on the world and tell them to work in plant science... -
Plasmids 101: Terminators and PolyA signals
TypeBlog PostPublishedMarch 31, 2016, 2:30 p.m....21439297. PubMed Central PMCID: PMC3622210. Schek N, Cooke C, Alwine JC. Definition of the upstream efficiency...(A) signals in combination with certain viral packaging systems has been associated with reduced viral... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m....References 1. Aurnhammer, C., Haase, M., Muether, N., Hausl, M.A., Rauschhuber, C.T., Huber, I., Nitschko...analyses needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting...had never worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit...transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV...intermediate into the single-stranded genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper...plasmid which encodes a transgene of interest that’s packaged into the virus. Two to five days after transfection...plasmids contain ITRs which are required for proper packaging of AAV, but also form secondary structures that... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog PostPublishedMay 15, 2014, 3:33 p.m.... of the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist...from quenching by aqueous solvent. This tightly-packed structure explains the importance of the entire...stability, GFP can be used in lineage tracking capacities in cell fate studies. It can also be used, when... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog PostPublishedAug. 9, 2016, 2:30 p.m....GENOME --local -f -k 10 --very-sensitive-local -L 9 -N 1 -U GRNA_23MERS -S GRNA_HITS.sam Many thanks to ...action at a distance.” Through the marvel of DNA compaction, an enhancer can increase the expression of a...millions of base pairs away. (For details on DNA compaction and the structural proteins that make it possible...details. DNA is a 3D Fractal Figure 2: DNA compaction. Graphics courtesy of beyondthedish (left) and...tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes per gram—and form...chromosomes we all know and love. To understand compaction, suppose you have a piece string that’s 10 meters..., are mere millimeters apart. Similarly, cells compact DNA in a way that brings linearly distant regions... -
Bioinformatics at Addgene
TypeBlog PostPublishedJuly 14, 2022, 1:15 p.m.... filtered the input channel to only use the first n items. To be sure, we went through much trial and ...complete package, all of the pieces of software that depend on each other for the entire package to run....in-house software has been crucial for us to keep pace with our current need to analyze nearly 300 sequences...sequencing data collection and analysis eventually outpaced our software, even with continuous updates. The... -
INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons
TypeBlog PostPublishedDec. 7, 2020, 2:53 p.m....M, Zhou Z, Smock DC, Tang K, Owens TK, Krishnappa N, Sachdeva R, Deutschbauer AM, Banfield JF, Doudna ...top): A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins...INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and technical considerations...INTEGRATE system follows a simple workflow: Clone spacer(s) into the CRISPR array Clone desired DNA cargo... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....information, you can fuse each FP fragment to the N- and C-terminal end of each PIP and empirically test...sequences, the amino acid sequences RSIAT and RPACKIPNDLKQKVMNH have been used successfully (Kerppola, 2006...expressed transiently by transfection or stably impacts the success of BiFC experiments. In either case...//doi.org/10.1101/pdb.prot076497 Kim Y, Jung JP, Pack C-G, Huh W-K (2018) Global analysis of protein homomerization... -
CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9
TypeBlog PostPublishedNov. 14, 2024, 2:15 p.m..... 2015;6. doi:10.3389/fpls.2015.01001 Ho TT, Zhou N, Huang J, et al. Targeting non-coding RNAs with the...start site function would also help to reveal the impacts of a lncRNA on nearby genes and its role in gene... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m....genomes. Cas9s from other species, like S. aureus and N. meningitidis require different PAM sites than SpCas9... 64 possible NNN PAM sequences adjacent to a protospacer. If ω-dCas9 could recognize a library PAM, it... -
AAVs in Retinal Gene Therapy
TypeBlog PostPublishedJuly 31, 2018, 12:55 p.m....Central PMCID: PMC4267879. 10. Samulski RJ, Muzyczka N., 2014. AAV-Mediated Gene Therapy for Research and...integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes. Unfortunately... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog PostPublishedJuly 2, 2020, 12:40 p.m.....2017.08.003 Liu, X., Chen, Y., Zhang, Y., Liu, Y., Liu, N., Botten, G.A., Cao, H., Orkin, S.H., Zhang, M.Q.,...that are distant linearly but close together in space. The chromatin “picture” is then developed by detecting... -
Antibodies 101: Introduction to Immunofluorescence
TypeBlog PostPublishedNov. 16, 2021, 2:15 p.m....https://doi.org/10.1007/978-1-4939-8935-5_26 Stuurman N. (2012) Introduction to Fluorescence Microscopy. iBiology...architecture of my cells,” or “How does this mutation impact the types of cells found in my tissue.” It is based...antibody performs differently and some antibodies impact the performance of other antibodies. But, once ...the right protocol, multiplex IF can give you impactful and beautiful results. Performing an immunofluorescence...fixation maintains sample morphology with minimal impact to the target epitope. There are multiple methods...exclusive to IF, but they can have a significant impact on the outcome of your IF experiment. Many of the...choose for your experiment will have a significant impact on the protocol you use and the ultimate outcome...