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...Addgene's Special Collections). Arguably, oncology has benefited most as cancer cells are amenable to culture...misidentified cell lines. The solution Fortunately it has never been easier to conduct high quality cell culture...identity. For example, perhaps a working cell bank has been made by a research institution (an excellent...at the scene of a crime (every human in the world has a unique STR profile) and can also be used as a paternity...topics. The LGC Group also provides standards and has laboratories to carry our independent quality control...

...firm The path to a career as a Grant Professional has been a long and winding path for many in the field...?  In the past 5-10 years, the Grants Profession has become just that, a profession. A grant professional...and the increasing regulatory process for grants has fostered the demand for experts in the field of grants... find your people Find someone who is working or has worked as a Grant Professional to mentor and coach...includes the social media community GrantChat. She has served in the nonprofit and government sector for...

... in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes...protein interactions in living cells. Since then FRET has also been used to measure conformational changes,...brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP. The stoichiometry...Fluorescence lifetime imaging microscopy method FLIM has been developed more recently and is the most rigorous...emergence of faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...

...policy. Since early March, the open access publisher has required authors to include a note as to where readers...how much data was necessary and reasonable. PLOS has since clarified their data sharing policy and recently...Who is to say that the initial data-generating lab has efficiently trawled their own data? A distinct (perhaps...many have multiple, complex panels. PLOS, itself, has not finalized its definition of necessary data requirements.... Secondly, science is highly technical and this has a natural, albeit unintentional, tendency to work...

... and polling, Blugene's new virus friend finally has a name! We thank you for your participation in suggesting...play on viral vector terms. And, the name Kendall has a special place in our hearts: our first two offices...

...Training as a scientist in the academic system has many pluses. I delighted in my graduate school years...call this building relationships because networking has gotten a bad reputation (as in…”I just hate networking...connections. A mistake made by many novice networkers has a potential to alienate connections. If meeting with...

... Science Guides Addgene has created the following guides to help you start learning...

...targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and variants ...scientific community. What are the features? CasPEDIA has collected relevant information on all the Cas enzymes...cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the other hand, generates...learn about Cas domains. CasPEDIA either directly has this information or links out to this content and...

...ElizabethKenkel) December 5, 2018 How social media has helped me in science It may have taken a bit of FOMO...Here are three examples of how using social media has helped me or my lab mates: For me, using social ...with social media as a scientist Using social media has enriched my professional development, and while it...helped you fit social media into your busy schedule. Has social media helped you advance your research or ...

...being targeted via RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly...sequence of the toehold switch. For example, Green has also developed toehold switches to detect Zika virus...protein motifs than previously thought. Sidhu’s group has made several hundred SH3 domain containing plasmids.... In addition to these constructs, the Sidhu lab has made hundreds of additional plasmids available through...

...and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...unsure of the next steps. Harvard PI Angela DePace has put personal development at the center of her lab... and postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine...

... field of induced pluripotent stem cells (iPSCs) has been around for 10 years. In that time, scientists... of Myc or other oncogenes after differentiation has been linked to tumor formation in iPSC-derived and...iPSCs compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared to MMLV-derived...mechanism between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...cell division. A drawback to this approach is it has lower levels of efficiency at generating iPSCs, usually...reprogram cells is that it’s simple and efficient. It has a high efficiency of reprogramming when compared ...

...wildtype Cas9 with optimized chimeric gRNA has high efficiency but has been shown to have off-target effects... the Genome Engineering Using CRISPR/Cas Systems has expanded out of control. To further consolidate these... a new system, developed by the Zhang lab, which has comparable efficiency to the optimized chimeric design...sequence is present in both alleles. The Zhang lab has found that if a CRISPR is used to engineer a locus...relative to where the homology ends? A9: The Zhang lab has always used homology arms that are less than 15 bp...recombination? A10: The most we've tried to insert so far has been 1 kb. We used homology arms that were 800 bp...for positive ones.  -- OR -- If your HR template has a selection marker such as puromycin, you can (also...

...objective performs essentially the same task but has different magnifying powers. When you look at the...

...to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts that can be assembled into...yeast MoClo is also for eukaryotes! The technology has been used to optimize protein secretion and expression...possible through MoClo. The MoClo Plant Parts Kit has everything you would expect in a MoClo kit plus the... general plant cloning experiments. Addgene also has two additional MoClo plant kits: Plant Parts Kit ...; Chamness et al., 2023). The Plant Parts II kit has a focus on infrastructure with shuttle vectors, several... gene silencing vectors. The Plant Parts III kit has a focus on transformation and genome engineering....Brychcy et al., 2023). For plant biologists, Addgene has not one, not two, but three CRISPR MoClo kits for...

... at the Georgetown University School of Medicine has used this gene editing system frequently over the...in human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor...via gene trap enrichment, whereas pAAV-TK-Acceptor has a conventional heterologous promoter-driven TK-neoR...editing compared to CRISPR editing CRISPR, of course, has started a revolution in gene editing. However, while... through aberrant double strand break repair, it has proven more challenging to develop high efficiency...

...reprogrammed less efficiently, no such correlation has been found between the age of the patients from whom...potentially promote tumorigenesis in vivo. The phenomenon has been recorded in past gene therapy trials and remains...reprogramming factors To date, the mRNA transfection method has not been used successfully to derive iPSCs from PBMCs...However, reprogramming of PBMCs by episomal vectors has yielded hiPSCs at a low efficiency (0.001-0.03%) ...efficiency: the pCXLE episomal vector toolkit It has been shown previously that suppression of TP53 enhances...

...quantified by cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-...performed on all new viral vector preparations. Addgene has used this method to generally validate our viral ... Material (AAV SFM), which is an AAV sample that has been quantified by 16 labs around the world and can...Hindson et al., 2011, Hindson et al., 2013), which has been shown to be more precise and more repeatable...2011): 8604-8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL,...

... maintain itself within host cells.  The plasmid has multiple ori that are functionally differentiated...various hosts (Pagatto and Dillon 2001).  The plasmid has its own replication initiation factors and proteins...the host. Host range depends on whether the host has the components to recognize the ori and allow for...restriction enzymes (Meyer et al. 1977). So if a plasmid has few restriction sites, it may be able to sneak past...commonly used bacteria in lab manipulations. Table 1 has a detailed list of the bacterial species these plasmids...

... 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and ...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room for additional...the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type counterpart...amino acids, can also be packaged into AAV. NmeCas9 has the added advantage that it can be turned off by ...daunting, it’s actually a challenge that the field has overcome with split AAVs. In general split AAVs break...