...attach the gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest...researchers as we could, and Addgene is a wonderful vehicle for us to do that.” At Addgene, we are equally excited...is available in an easily searchable format on Addgene’s Michael Davidson Fluorescent Protein Collection...collection with over 300 backbones alone,” said Addgene’s Lianna Swanson, who has been working with members...the scientific community," says Joanne Kamens, Addgene's Executive Director, "and to promote the great ...

...inducible gene-expression sequences, and more. By combining these components, one can generate constructs...localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu... might not, however, be appropriate for cloning genes from organisms with high CpG dinucleotide representation...to combine the functions of different genetic elements to generate non-natural tools such as novel biological...Resources on Addgene.org Browse our Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid...nutshell, is the systematic combination of different genetic sequences into plasmid DNA. In high throughput ...techniques, although the specific sequences of the genetic elements may differ (e.g., a set of various mammalian...

...nucleus AAVs CREATEd for gene delivery to the CNS and PNS Resources on Addgene.org Find the Spaghetti Monster...with the development of methodologies to deliver genes to the CNS have recently made it much easier to ..., and the long-term expression of AAV delivered genes. AAV serotypes have been extensively studied in ...essential for neuroscience research. The use of genetic tools for visualizing and perturbing circuits together...Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the brain because of its low cytotoxicity...connections to that population. Limitations of common genetically encoded neuronal tracers Neuronal tracers such...influenza hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus 5-derived epitope (V5), the...

...-in mutations and gene editing Targeted genome editing events fall into two general classes: knock-in ...HDR. If your Cas9 cuts in the coding region of a gene, make sure that the PAM edit you introduce is a ...Guo, Y., Tian, Qigi, Lan, Y., et. al. An efficient gene knock-in strategy using 5’-modified dsDNA donors...translation by generating a frameshift mutation. The majority of cellular repair events will generate these types...family member which generates sticky ends upon cutting. Cas12a may promote HDR by generating ssDNA overhangs...Taking the road less traveled and generating a knock-in cell line instead of a knock-out? We’ve got you...cells, but a less frequent DNA repair mechanism in general. On average, it accounts for less than 10% of DNA...

... (Shmakov et al., 2015). They were using Cas1, a gene commonly associated with CRISPR arrays and involved...analysis, they identified 53 potential candidate genes that fell into 3 categories based on the architecture...expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although the...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check Out Our CRISPR Featured...27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an exceptional genome engineering... PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky, Alexandra, et al. "Two distinct...PMID: 27669025. Find plasmids from this paper at Addgene.  3. Gootenberg, Jonathan S., et al. "Nucleic ...

...directional gene transfer, analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer...from this publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex Gene Editing by CRISPR–Cpf1 Using...26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide View...Again, they were in for a surprise! Whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1...27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik, et al. Genome-wide analysis reveals...27272384 Find plasmids from this publication at Addgene. Kleinstiver, Benjamin P., et al. "Genome-wide...27347757. Find plasmids from this publication at Addgene. Makarova, Kira S., et al. "An updated evolutionary...

...Many neuroscience experiments that require gene expression in a specific cell type rely on transgenic... (recognized by Flp), to either flip or excise a gene that is located between them. Early recombinase-...Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678. https://doi.org/10.1089/hum.2019.277...Peripheral Nervous System and Relevance to Human Gene Therapy. Front Mol Neurosci 13. https://doi.org/...end of the transgene. Since loxp and lox2272 can not recombine, this ensures that the transgene is no longer...flank a transgene (homologous pairs) or used different scrambled sequences to flank the transgenes (non-... express Cre or Flp. Here at Addgene we quantify the amount of transgene recombination for every lot of...

...experiment. PITChing MMEJ as an Alternative Route for Gene Editing Michohomology-mediated end joining (MMEJ...template to increase efficiency of insertion and gene replacement. Care to share your favorite CRISPR ...Blugene and I represented Addgene at the recent Keystone meeting on Precision Genome Editing with Programmable...hopes to evaluate genome editing pipelines and generate a common lexicon for genome editing studies. These...desired edit, and I want to make sure that I (and Addgene) don’t forget this in the future. Here’s a list... to keep current with the latest and greatest! Addgene resources 3 Tips to Improve HDR Efficiency for ...

...with genes central to the replication process removed. This means the vector can deliver a genetic payload...vectors to reproduce when experimentally necessary, genes for the required proteins are packaged in separate... More resources from Addgene The AAV Data Hub Addgene's Viral Services Addgene's Viral Protocols...viruses bud out, instead of lysing, but they do generally still kill the infected cell.) This is in contrast...labs. Viral vectors are viruses that have been genetically modified to either limit or completely eliminate...10.1007/978-3-319-07758-1_2 More resources on the Addgene blog Viral Vectors 101: Viral Applications Viral...

...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many ...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520... exhaustive refinement, it was not possible to generate an optimal Rluc8-Gα construct for two of the 16...s0014-5793(02)02775-8 Additional resources on the Addgene blog Browse all of our fluorescent protein blog...Find other fluorescent biosensors Resources on Addgene.org Browse the catalog of fluorescent protein plasmids...