...expression and knockdown. Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P. Nat Methods. 2006. Feb;3(2...

...post. There’s so many great tips for getting the most from your conference experience so we unfortunately...them all. What are your top tips for getting the most out of your scientific conference? Resources on...

.... What aspect of GeoCas9 are you most excited about? Thermostability? Longevity in human plasma? Something...cocaine esterase engineered to have increased thermostability also had a longer lifetime in human blood. ...lower than optimal temperatures. GeoCas9’s thermostability could also be used with in vitro molecular ...CRISPRing! References Harrington, Lucas, et al. "A thermostable Cas9 with increased lifetime in human plasma... thermostable cas9...

...reproducibility. Read on for some tips for getting the most out of your lentiviral transduction experiments....point to consider is how the virus is being titered. Most titering approaches transduce into highly permissive...

.... Our most popular kit of the year was the Golden Gate TALEN Kit, which continues to be our most requested...distributed through Addgene over our first 10 years. 10 most distributed plasmid technologies pSV-PGC1 - Ok,...Fire's set of vectors for C. elegans research was our most requested item of 2005. shRNA - In 2006, we saw ...two plasmids from Didier Trono's lab are our two most popular plasmids of the past 10 years. Induced Pluripotent... Yamanaka's lab and James Thomson's lab were the most frequently requested. Ubiquitin - One particular...interest in from 2008 to 2009 was ubiquitin. The most requested ubiquitin plasmids were pRK5-HA-Ubiquitin-WT...

... other Addgene news Find the most recent plasmid blog posts Find the most recent CRISPR blog posts Resources...

...wanted to celebrate our depositing PI’s who have the most highly requested plasmids. Currently, there are ...

... the Quake Lab. Identifying IgnaviCas9, a hyperthermostable Cas9 This year it was wonderful to learn that...Ignavibacteriae phylum genome. IgnaviCas9 is a hyperthermostable Cas9 CRISPR nuclease that is active up to ...optimal sgRNA spacer length of 22-25 nts, and is thermostable between 31-100°C. They published their finding... in a bioRxiv article and in PNAS. Two other thermostable Cas9 enzymes have previously been characterized...ThermoCas9, IgnaviCas9 is a type II-C Cas9. IgnaviCas9 is most similar to a type II-C Cas9 from Parvibaculum lavamentivorans...What can you do with IgnaviCas9? Why would a thermostable Cas9 be useful? It can enable applications where...steps in the protocol. IgnaviCas9’s incredible thermostability makes the enzyme particularly useful. Additional...

...yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic chromosomes, and ...

...recommend it except in the most dire of circumstances.) Multiplex assays Mo’ proteins, mo’ problems may not be...region, where an antibody binds to its target. While most secondary antibodies recognize broad isotype classes...across applications. Western blots, for instance, almost always use the indirect approach, while flow cytometry...assay and protein of interest is often one of the most challenging components of any immunoimaging assay...incorrect. With the exception of flow cytometry, most immunoimaging assays do not allow for absolute quantitative... and occasionally ask themselves and others that most perplexing of questions: “who moved my goat anti-rabbit...

...immune system’s response to foreign bodies, are the most commonly used affinity reagents. Antibodies are ...against the antigen, packaged, and sold. This is the most cost-efficient way to produce antibodies, with the... IgA2 IgD Coexpressed with IgM on the surface of most immature B cells; function is unknown Monomer with...anaphylactic shock Monomer with a valency of 2 IgG The most abundant isotype in blood, also found in tissues...Produced during the primary immune response; found mostly in blood Pentamer with a valency of 10 Antigen ... for exact storage requirements. As a guideline, most antibodies can be stored at 4 °C for short-term ...allow detection of the antibody:protein complex. The most commonly used types of signaling molecules are horseradish...

... and adaptability, CRISPR has rapidly become the most popular genome engineering approach. Engineered ...ease of generating gRNAs makes CRISPR one of the most scalable genome editing technologies. This advantage...repair (HDR) pathway The NHEJ repair pathway is the most active repair mechanism, and it frequently causes... information, jump to Plan Your Experiment ). In most cases, NHEJ gives rise to small indels in the target...using conjugation and stably integrated into the chromosome. Browse Plasmids: Activate , Repress/Interfere...and synthesis, as well as the ability to target almost any genomic locus, make CRISPR the ideal genome... viability, and tumor metastasis. Currently, the most popular method for conducting genome-wide screens...

...CRISPR/Cas9 in Caenorhabditis elegans. Mos1 Based Transgenesis miniMos Transgenesis Kit - Erik Jorgensen Lab... related miniMos plasmids can be found on the Jorgensen Lab page. More information on miniMos and MosSCI...

...dollars spent each year. Unfortunately, the causes of most neurodegenerative diseases are only partly understood...no effective therapies to prevent, treat, or cure most of these disorders. An integrated study of the overlapping...introduction to the genetics associated with some of the most common disorders followed by a list of some of the...ten million people worldwide are living with PD, mostly people over the age of 60. There is presently no...Alzheimer’s Disease Alzheimer’s disease (AD) is the most common cause of dementia. Symptoms include memory...

...repetitive and non-repetitive genomic loci, as well as chromosome painting in living cells. Visualizing a specific...labeled proteins to the given region. For example, chromosome-specific repetitive loci can be efficiently visualized...co-delivering multiple gRNAs that tile the locus. Chromosome painting requires delivery of hundreds of gRNAs...gRNAs with target sites throughout the chromosome. Mammalian Plasmid Gene/Insert Promoter Selectable Marker...

... guides around the region to find the most efficient guide (most guides would work, but they sometimes...multiplexing CRISPR to target multiple genome loci, the most efficient and easiest way is to co-transfect several...homology arms be for efficient recombination? The most we've tried to insert so far has been 1kb. We used...

...addresses", where dCas9 is going to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression...Preprint at bioRxiv Detailed information about chromosomal integration of pLC97 can be found in this article...article: St-Pierre et al., One-Step Cloning and Chromosomal Integration of DNA, ACS Synthetic Biology, 2013...

...localization) the C - t erminal D omain (CTD) The most common missense mutations cluster in the MBD and...R168X, R255X, R270X, R294X, and R306C) account for almost 46% of all Rett syndrome cases. Research Milestones... and the direction and degree of skewing of X-chromosome inactivation. J Med Genet . 44, 148–152. (Link...

...Parkinson's Michael J Fox Foundation MJFF 40386 pBACgus_Sumostar/LRRK2 1867-2186 LRRK2 His, TEV polH Parkinson's...Parkinson's Michael J Fox Foundation MJFF 40387 pBACgus_Sumostar/LRRK2 1867-2186_G2019S LRRK2 His, TEV polH...Parkinson's Michael J Fox Foundation MJFF 40388 pBACgus_Sumostar/LRRK2 1867-2186_C2024S/C2025S LRRK2 His, TEV...Parkinson's Michael J Fox Foundation MJFF 40389 pBACgus_Sumostar/LRRK2 1867-2186_T2031E LRRK2 His, TEV polH...Parkinson's Michael J Fox Foundation MJFF 40390 pBACgus_Sumostar/LRRK2 1867-2141 LRRK2 His, TEV polH Parkinson's...Parkinson's Michael J Fox Foundation MJFF 40391 pBACgus_Sumostar/LRRK2 1867-2161 LRRK2 His, TEV polH Parkinson's...Parkinson's Michael J Fox Foundation MJFF 40392 pBACgus_Sumostar/LRRK2 1867-2176 LRRK2 His, TEV polH Parkinson's...

...presence of specific molecules or other factors. Most often, a conformational change in the sensing region... astrocyte calcium signaling in the hippocampal mossy fiber pathway. Neuron. 2014 Apr 16;82(2):413-29....from ATP-Mg Hydrolysis Contributes to Mitotic Chromosome Condensation. Curr Biol. 2018 Feb 5;28(3):444...intracellular environment of S. cerevisiae . The MOSAIC Kit from the Cohen laboratory includes 19 lentiviral...