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...the Liu lab discovered that cellular mismatch repair contributed to a large number of unintended prime ...stability Traditional sgRNAs are protected from cellular degradation by Cas9 binding, but pegRNAs have ...pegRNAs, but combining PE7 with epegRNAs resulted in similar or reduced editing efficiencies, likely because...edits to opposite strands of the DNA and skip the cellular mismatch repair step (Anzalone et al., 2022) PASSIGE...combine prime editing with a recombinase to install large sequences (up to 10 kb) into the genome (Pandey ...deletion, replacement, integration and inversion of large DNA sequences with twin prime editing. Nature Biotechnology...Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22...

...are not limited to, ones targeting muscular dystrophy, cardiovascular disease, inflammatory diseases, blindness...TTR is primarily made in the liver. Similarly, for cardiovascular disease approaches targeting cholesterol...et al). CRISPR engineering of CAR-T cells is particularly promising because of its potential for allogenic... polymorphisms (SNPs) in the TTR gene and is molecularly characterized by misfolding of the TTR protein...mutations or introducing a desired edit, but this particular class of treatments has quite the opposite aim...therapy currently in clinical trials is somewhat similar (Hussein et al). It employs Cas9 to cut at two ...two sites within the viral genome to excise a large portion of it. This ongoing trial is sponsored by Excision...

...receptors The receptor you choose is largely dependent on what cellular response you aim to manipulate. It...pathways, giving rise to the same cellular responses. The cellular response set off by a DREADD depends ...normal brain function. Chemogenetics has become a popular tool in the field as it provides a noninvasive ... where specific tissues, cell types, or even subcellular regions of a neuron need to be targeted. Targeting...nonnative ligands. However, they retain their intracellular interactions and still set off the same pathways...been derived from muscarinic receptors. The most popular agonist for muscarinic receptor derived DREADDs...such as the serotonin 3 receptor, will promote depolarization in neurons. Alternatively, combining with the...

...Viral delivery of TnpB for plant genome editing LARRYv2 pooled libraries for cell barcoding Addgene’s own...deposited a variety of constructs for visualizing cellular components (nucleus, endosomes, lysosome, golgi...found itself in the spotlight. TnpBs are gaining popularity, especially among plant biologists, due to their... (learn more in our recent blog post on TnpB). Similar to CRISPR-Cas, TnpB is directed to genomic targets...virus, to deliver a TnpB (named ISYmu1) to the popular plant model Arabidopsis (Weiss et al., 2025). By.../s41477-025-01989-9.   Make a splash with new LARRY barcoding libraries By Mike Lacy Whether you’re ...method is their updated Lineage and RNA Recovery (LARRYv2) barcoding libraries. The team used these libraries...

.... An accidental release would be particularly dangerous for molecular biology and genetics because the...such as malaria and dengue for many years. Since more than a thousand children died of malaria today, ...safeguards and confinement strategies are available? Molecular confinement involves building gene drives that... tropical mosquito vectors of diseases such as malaria and dengue are currently being performed in regions...performed with two or more confinement methods. Since molecular, ecological, and reproductive confinement typically...cost of an accidental release. Science relies on popular support, which in turn depends on widespread trust...shared environment, they must never be used without popular support2. An accidental release could delay real-world...

... as to make plants, and agricultural crops in particular, resistant to disease. They also devise biosensors...biosensors designed to elucidate the molecular interactions that go on between plants and their pathogens...biology Addgene: Tell me about the Golden Gate Modular Cloning (MoClo) Kit. What does it include? Patron...to show that they work well in plants and give similar stability to transcripts as the ones that people...we are collecting sequences that will provide a similar resource for those involved in monocot engineering...workhorse. Tips & tricks Addgene: Do you have any particular tips for those looking to get started with MoClo... parts. That was the motivation for releasing a large parts kit. Once you have so many parts available...

...lentivirus). Large cargos, such as the ~11 kB cDNA for dystrophin, the gene mutated in muscular dystrophy... being inserted into new target sites. This is similar to how retroviruses, like HIV, replicate. Class...can be inserted between the transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a...vertebrates and integrates in human cells at rates similar to retroviral vectors. The hyperactive version ...piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase...with a maximum cargo capacity of ~200 kB of DNA. Similar to piggyBac, Tol2 also prefers to integrate at ...transposons Now that you’ve learned about some of the popular transposon systems, let’s take a look at how they...

... etc. Contamination by humans accounts for the largest source among those mentioned above. The contaminants...and then specific enzymes are used which trigger cellular mechanisms that generate luminescence. This can... in the form of small clusters, apart from the cellular DNA. Fluorescent DNA staining is another DNA staining...mycoplasmas, contamination by other bacteria or extracellular fluorescent signals produced by these bacteria...masks. Make sure to change the gloves and masks regularly instead of keeping them in the pockets of your...preventing mycoplasma growth. Also, prolonged use of a particular antibiotic can induce resistance for other bacteria...cross-contamination. CO2 incubators should be cleaned regularly and dampness of any kind should be avoided. Distilled...

...Resources on Addgene.org Basic Molecular Biology Protocols Protocol Videos Molecular Biology Reference Pages  ...T4 DNA ligase) are critical reagents for many molecular biology protocols, including vector-insert joining...sequencing (NGS) library construction, and circularization of dsDNA (6). Less commonly utilized in vitro...are sensitive to distortions in helix shape (22). Large purine:purine mismatches and most smaller pyrimidine...with products identifiable and quantifiable by capillary electrophoresis (CE) (see below). This method ...novel 16S rRNA gene PCR-ligase detection reaction-capillary electrophoresis assay." Journal of clinical microbiology.../LDR/flow-through hybridization assay using a capillary tube, probe DNA-immobilized magnetic beads and...

...colonies that are comprised of cells with a large nucleus, large nucleoli, and little cytoplasm. Positive...oncogenes, such as Myc. Viral vectors also have the largest genomic footprint of iPSC generating methods due...infect dividing and nondividing cells and have a larger cloning capacity (8-10 kb) than other retroviral...factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared to MMLV-derived...transfection efficiency is cell-type dependent and larger plasmids have lower rates of transfection. Minicircles...other methods. This could be a reflection of the large size of the plasmid or silencing of the plasmid ...modifications are used to protect the RNAs from cellular defenses against ssRNA: phosphatase treatment,...