...Intuitively, one would expect that assembling relatively small, repeat-laden DNA regions into a single construct...sequence-specific TAL effectors for modulating mammalian transcription. Nat Biotechnol. 2011 Feb;29(2):...

...into the host genome. AAVs are limited by their small packaging capacity. Adenovirus Plasmids | Guide ...delivery of genetic information to hard-to-transfect mammalian cells. In addition, "viral pseudotyping" can be...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish... C->T (or G->A on the opposite strand) within a small editing window specified by the gRNA. Adenine base...

...properties of the final product, particularly in large animal studies. Endotoxin contamination is minimized by...MRP 30 kDa subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma...

...gene encoded by the transfer plasmid is toxic to mammalian cells, it can be difficult to efficiently produce...gateway destination plasmid for cDNA expression Ramalho-Santos 24129 pULTRA 3rd bi-cistronic expression...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish...tagging of proteins expressed from their natural chromosomal context. These CRISPR tagging methods allow for...

...Flp, here expressed through a double transgenic animal strategy (PV-2a-Cre; SOM-IRES-Flp). Image from ...combinations of transgenic recombinase-expressing animal lines and retro-grade expressing viruses delivering...

...fluorescent proteins. The majority of the ORFs are of mammalian origin and include localization data found below...

...Jonathan Katz Nathan Lawson Kathrin Plath Miguel Ramalho-Santos Derrick Rossi Michael Ruppert Michel Sadelain...

...Dana-Farber Cancer Institute Finding the Causes of Small Intestinal Neuroendocrine Tumors Ramesh Shivdasani...

...Collection Protein Expression Cell Migration Consortium Malate Dehydrogenase CUREs Community (MCC) Collection ...

...229E) Expression type (e.g. bacterial, yeast, or mammalian) Tags (e.g. 6xHis, Flag, StrepII, HA, Myc, SUMO...

...expression plasmids (AsCpf1 and LbCpf1) for use in mammalian cells SpCas9 variants with no detectable off-target...

...pAquaN1 Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a ...compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial...pCEP4YPet-MAMM Yellow Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet...Yellow Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses...N-terminus of monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus...C-terminus of monomeric Cerulean mTurquoise2 Cyan Mammalian Constructs to target mTurquoise2 to various subcellular...Expresses CyPet with C-terminal His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus...

...cancer development. These pathways are important to normal cell signaling and physiology, and, when dysregulated...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish...

... yeast two-hybrid assay are not optimal for studying native mammalian protein interactions, whereas others...is needed to assess its function in mammalian cells or other animal models, but this technique has laid...p2A-ferritin-p2A-mCherry Mammalian Expression 74308 pcDNA3.0-Magneto2.0-p2A-mCherry Mammalian Expression 74334...sheared genomic libraries are cloned downstream of a minimal promoter sequence. Should one of these random stretches...it will activate its own transcription from the minimal promoter and its strength can be determined from...RAS Pathway Clone Collection 2.0 Ras proteins are small GTPases involved in cell signaling pathways that...been difficult to target therapeutically due in no small part to the central role Ras proteins play in cell...

...expression in neuronal cells. U6 small RNA expression shRNA From the human U6 small nuclear promoter Constitutive...expression) you need to use an RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP...Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc) require unique promoters ...leakiness" of transcription may occur. This is normally not a big issue for cells, but it may confound...listing some of the most common bacterial and mammalian promoters. These lists is by no means exhaustive...considerations CMV General expression mRNA Strong mammalian expression promoter from the human cytomegalovirus... types. EF1a General expression mRNA Strong mammalian expression from human elongation factor 1 alpha...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish...

... by the heat. Isolating male and female flies Differentiating male and female flies is essential for setting...of their abdomens, males have black coloration whereas females do not. Also, males have sex combs on their...is important to isolate females from males while they are still virgins. Females begin mating ~6 hours ...trying to get males to recombine two mutations onto the same chromosome. You should use females for that. ...their forelegs which are black, bristly hairs. Females do not have sex combs. To make genetic crosses ... bodies. This is the ideal stage to select for females because you can be certain that they are too young...fourth chromosomes, you are a very unlucky person. Animals are diploid except for their germ cells (sets of...

...screening 768 F1 animals). Recovering rare editing events in a sea of unedited animals struck me as a “...from 11-100% of F1 animals carrying precise knock-ins, and homozygous knock-in animals can be obtained in... in eight days. The only animals on a plate, other than the parental animal, are rescued progeny, which...for any selective marker to remain in the edited animal. Development of CRISPR/Cas9 editing strategies...must be removed, which can be done by isolating animals with particular visible phenotypes. These approaches...conditional-lethal mutation to identify edits with minimal screening effort. Selecting for repair of a lethal...unedited “haystack”, facilitating recovery of edited animals. Figure 1: In the background are 504 restriction...

...difficult-to-target antigens that are intractable via animal immunization. Addgene's antibody catalog is a community...

...difficult-to-target antigens that are intractable via animal immunization. IPI is a nonprofit research organization...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish...

...website contains additional information CRISPR Optimal Target Finder Identifies gRNA target sequences ...

...pAAV-FLEX-EGFPL10a EF1a-driven and Cre-dependent EGFP-tagged ribosomal L10a 5, rg* Heintz , Nectow , Schmidt Neurophysiology...

...scientist Twitter users are male and 40% female, for a ratio of female to male = 0.629 (the closer this ...closer males and females are to parity - the smaller the number, the more it leans towards males). This...dataset. 3. 60% of scientists on Twitter are male and 40% are female Figure 2. Gender Ratio of Scientists on...is more skewed towards men than the ratio of female to male Internet users that use Twitter (0.875), but...but less skewed than the ratio of female to male scientific authorship in the United States (0.428). 4...

...induced by the addition of a small molecule, and protein levels can return to normal once the drug has been ... expressed beyond the tag and small molecule. It does work in mammalian cells, mice, and even yeast, though...conditions (typically the addition of an exogenous small molecule). Degron tags can range in size from several...degrons and some of the nuances to each one.   SMASh Small molecule-assisted shutoff (SMASh) functions on the...itself when you want the protein around, leaving a minimal additional sequence (7 kDa). This system also requires...are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation tag (dTAG) system...selective degrader. The novel degrader, dTAG13, is a small molecule which induces dimerization of the FKPB12F36V...

... but it’s really just formaldehyde. Formalin: You may also come across formalin, which refers to a saturated...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative...saturated solution of formaldehyde (saturated being 37%). Formalin solutions often contain methanol as ...order to slow the polymerization of formaldehyde.  Note: Formaldehyde in solution will naturally polymerize... maintaining cellular and tissue morphology. Formaldehyde is the most commonly used cross-linker for IF...fixative. PFA is a polymer of formaldehyde that, on its own, is not a fixative. However, when PFA is dissolved...solution and heated, it depolymerizes, producing formaldehyde. Often people will call the resulting solution...

...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish...