...We’re excited to welcome Karen Katz, Esq and Carl Paratore, MBA, CPA to the Addgene Board of Directors... Daisy, or out on golf course! Learn more about Karen Katz, Esq Ms. Katz is the Director of Intellectual...Property Programs at Suffolk University Law School. Karen graduated with highest honors from the University...associated with Hale & Dorr and Crowell & Morning. Karen was a founding member of Mass Medical Angels, a ...

...Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar S, Davies BW, Ellington AD, Moran NA, Barrick JE ...   This system is distinct from those used in eukaryotes in that CRISPR isn’t the primary editing force... similarly to CRISPR/Cas9 systems adapted for eukaryotes in that Cas9-mediated cleavage induces HDR to...Marraffini lab used a native minimal CRISPR array (Bikard et al., 2013), the Qi lab employed a gRNA-based...design more familiar to those using CRISPR in eukaryotes (Qi et al., 2013). As in other systems, catalytically...efforts creating tools for CRISPRi and CRISPRa (Bikard et al., 2013). By targeting either a catalytically...Phagemids (Selle et al., 2020, Citorik et al., 2015, Bikard et al., 2014). When the CRISPR system targets these...

... longevity in eukaryotes and promote transcript degradation in prokaryotes.  Prokaryotic termination   ...broken down into two broad categories, prokaryotic or eukaryotic, based on the functional elements they...they contain. Plasmid DNA in both prokaryotic and eukaryotic systems must be transcribed into RNA, which...characterized a set of prokaryotic terminators and deposited several with Addgene (1). Prokaryotic polyadenylation...Although mostly thought of as a eukaryotic-specific process, prokaryotes also add poly(A) tails to certain...of mis-folded RNAs. Eukaryotic termination and polyadenylation Unlike prokaryotes that have a single RNA...not as well studied as prokaryotic termination, the basic processes for eukaryotic termination are understood...

..., F. E., Moeller, L., Kocalar, S., Oshiro, R., Makarova, K. S., Macrae, R. K., Koonin, E. V., & Zhang,...doi.org/10.1126/science.adi1910 Chylinski, K., Makarova, K. S., Charpentier, E., & Koonin, E. V. (2014...j.molcel.2023.12.034 Kolesnik, M. V., Fedorova, I., Karneyeva, K. A., Artamonova, D. N., & Severinov, K. V. ...CRISPR-CAS systems: deciphering the most complex prokaryotic immune system. Biochemistry (Moscow), 86(10),...doi.org/10.1134/s0006297921100114 Koonin, E. V., & Makarova, K. S. (2019). Origins and evolution of CRISPR-Cas.../doi.org/10.1098/rstb.2018.0087 Koonin, E. V., Makarova, K. S., & Zhang, F. (2017). Diversity, classification...–78. https://doi.org/10.1016/j.mib.2017.05.008 Makarova, K. S., Wolf, Y. I., Alkhnbashi, O. S., Costa,...

...tightly regulated. Common promoters for eukaryotes and prokaryotes We have included two reference tables...bacteria this is done by a single enzyme; however, eukaryotes have muliple polymerases which are each responsible...subset of RNAs. To gain this specificity, the eukaryotic RNAP can recognize and bind to specific promoter...similarly variable. Bacterial promoters only work in prokaryotic cells and typically only in the same or closely...which they were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc) require...and complex, having fewer parts than those in eukaryotic cells. Some promoters are constitutively active...start when trying to pick your perfect promoter! Eukaryotic Promoters   Promoter   Primarily used for ...

...year, scientists hijacked survival tactics of prokaryotes to deliver the technological biological blockbuster...demands of the biotech field, let's look at how prokaryotes originally utilized CRISPR/Cas as a powerful ...these studies led scientists to hypothesize that prokaryotes developed an adaptive immune system that utilized... of bacterial genomes utilize this system (4). Makarova et al. grouped the systems into three distinct...system seems like the winning ticket to ensure prokaryotic survival; however, viruses, too, are famous for...dynamic range of CRISPR systems between and within prokaryotic species suggest the presence of a variety of ...an interesting "arms race" between phages and prokaryotes; the ultimate victor will be the species who ...

...methods which can be used in prokaryotes, transfection is only done in eukaryotic cells. Transfection is the...introduction of foreign DNA or RNA into bacteria or eukaryotic cells is a common technique in molecular biology...foreign DNA or RNA is introduced into bacterial or eukaryotic cells via a virus or viral vector. One example...use transduction to introduce foreign DNA into eukaryotic cells, like mammalian cell lines. This can be...foreign DNA is deliberately introduced into a eukaryotic cell through non-viral methods including both...and viruses. References Baltrus, David A., and Karen Guillemin. "Multiple phases of competence occur ...to Genetic Analysis. 7th edition (2000). Meibom, Karin L., et al. "Chitin induces natural competence in...

...a versatile tool developed for use in prokaryotes and eukaryotes, works via direct activation. In this...promoter activation. One of the most commonly used prokaryotic promoters is the negative inducible pLac promoter...Negative inducible promoter pBad is another popular prokaryotic promoter often used for bacterial protein purification...similarly variable. Bacterial promoters only work in prokaryotic cells and typically only in the same or closely...which they were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc.) require...inducible promoter not be too leaky. For inducible prokaryotic promoters, pLac is known to be slightly leaky...

...one form of instability. Prokaryotes can also have a hard time folding eukaryotic proteins. You can get ...improve solulibility and folding of eukaryotic proteins in prokaryotes; single step purification with amylose...make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes to degrade... Not great for purification; too cleavable in eukaryotes TAP GRRIPGLINP WKRRWKKNFI AVSAANRFKK ISSSGALDYD...recognition site. In fact, SUMO cannot be used in eukaryotes because there is already too much SUMO protease...

...transposons are in the genomes of many other prokaryotes and eukaryotes, and make up ~45% of the human genome...genetic elements that are found in both prokaryotic and eukaryotic genomes.  Components of a transposon ...another tool to move DNA around or into eukaryotic and prokaryotic genomes. They require just two key components...EJ (2007) DNA Transposons and the Evolution of Eukaryotic Genomes. Annu Rev Genet 41:331–368. https://doi.org...

...Ptashne, M. A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor....1980s saw a flurry of discovery in the field of eukaryotic transcriptional activation and cell biology. ...protein degradation, access to the nucleus in eukaryotic screens and fusion configuration. The possibility...order to bind DNA and activate a reporter gene in eukaryotic cells, fusion proteins must gain access to the...from the transcription-activating function of a eukaryotic regulatory protein. Science. 1986; 231:699-704...

...aminoglycosides, including G418 which is commonly used for eukaryotic antibiotic selection. Essentially, this gene ...gene is a 2-in-1 providing both bacterial and eukaryotic selection, whereas other plasmids must carry both...is an all-in-one antibiotic — it is usable in eukaryotic cell culture, yeast, bacteria, and even plants...colonies Kanamycin Excellent No Bacteria and eukaryotes (G418) {% icon icon_set="fontawesome-5.14.0" ...} Zeocin Excellent No Bacteria, plants, and eukaryotes Not Tn5-containing cells {% icon icon_set="fontawesome...

...expression systems have been improved for both eukaryotic and prokaryotic expression in recent years, with many...cell-free expression of proteins in both prokaryotic and eukaryotic cell extracts (Gagoski et al 2015). He...sequence bypasses the 5′ mRNA cap required for eukaryotic systems and instead directly engages the ribosome...

...guest blogger Kartik Lakshmi Rallapalli from the University of California, San Diego. Kartik Lakshmi Rallapalli...This post was contributed by Kartik Lakshmi Rallapalli, a graduate student at the University of California...our understanding of the base editing enzymes. Kartik’s research focuses on the use of molecular dynamics...the realms of art and science. Follow her at @kartik_lakshmi. Additional resources on the Addgene blog ...

... from Stanford University, led by neurobiologist Karl Deisseroth, combined optical and genetic methods...30503173. PubMed Central PMCID: PMC6317997. Deisseroth, Karl, et al. "Next-generation optical technologies for...17035522. Pubmed Central PMCID: PMC2820367. Deisseroth, Karl. "Optogenetics." Nature methods 8.1 (2011): 26. ...PMCID: PMC3979449. Fenno, Lief, Ofer Yizhar, and Karl Deisseroth. "The development and application of ...photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas." Nature 311.5988 (1984): 756. PubMed...

...We all know that yeast is a powerful eukaryotic model organism – its genome is easy to manipulate, it...Yeast as a model organism Yeast are single cell, eukaryotic organisms formally classified as fungi. They ... Amenable to genetic manipulation The first eukaryote (S. cerevisiae) to have its genome fully sequenced...specific oxidative reactions. They are found in all eukaryotic cells and peroxisomes are where methanol metabolism...

...applications. Find a chromoprotein for your experiment! Eukaryotic chromoproteins for bacterial synthetic biology...common chromoproteins used in the lab come from eukaryotes, they can be difficult to directly use in bacteria.... https://doi.org/10.1007/bf00428249 Chan MCY, Karasawa S, Mizuno H, Bosanac I, Ho D, Privé GG, Miyawaki...Forster, A. C. (2018). Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology...

...described below. IRES Elements Translation in eukaryotes usually begins at the 5’ cap so that only a single...it promotes translation of the viral genome in eukaryotic cells.1,2 Since then, a variety of IRES sequences...employed by scientists. 2A cleavage is universal in eukaryotic cells, and, although some scientists report close...References: 1. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus...

...Many thanks to our guest blogger, Aneesh Karve! Aneesh Karve is CTO at Quilt Data. Quilt is a collaborative...This post was contributed by guest blogger, Aneesh Karve, CTO at Quilt Data. This post was originally published...for genomics. In this article, Quilt CTO Aneesh Karve, shows how to design experiments that work anywhere...

... like C. difficile. In 2014, Citorik et al. and Bikard et al. developed sequence-specific, antimicrobial...phagemids were cytotoxic only for resistant bacteria. Bikard et al. used a phagemid system to target virulent... the phagemid or received a defective phagemid; Bikard et al. did not observe any mutations at the target... Find plasmids from this paper at Addgene. 2. Bikard, David, et al. “Exploiting CRISPR-Cas nucleases...