...vectors, recombination often occurs between the long-terminal repeats (LTRs). After recombination, one resulting...intramolecular DNA recombination. This naturally occurring process is mediated by recombinases like RecA and...undergo recombination, two smaller plasmids are formed. Viral vectors are prone to recombination In lentiviral...replicate.   How can I prevent recombination in my samples? Plasmid recombination can affect viral vectors ...have been engineered to further reduce recombination. Recombinase-deficient strains like Stbl2, Stbl3, ... repeating elements. Knowing how to amplify recombination-prone plasmids and testing your prep can save..., and NEB stable E. coli reduce the rate of recombination in samples. For a more detailed comparison of...

...to avoid MMR: 1) E. coli with inactivated MMR: Using E. coli with inactive MMR is definitely the easier...minutes quickly inactivates the repressor and allows for expression of the recombination genes.  After ...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...genome engineering and is based on homologous recombination. It allows for direct modification of DNA within...called recombineering - short for homologous recombination-mediated genetic engineering.  It can be used... These enzymes then catalyze the homologous recombination of the substrate with the target DNA sequence...nucleotides of homology to the target site for recombination. Key components of the Lambda Red recombineering...

... you encounter mycoplasma contamination. Causes of mycoplasma contamination The mycoplasmas enter the ...above. The contaminants can spread through dirty clothing, lab wears, human speech near the laminar airflow... risk of contamination. Mycoplasmas can spread from these sources through cross-contamination and due ... a source of mycoplasma contamination. Due to its turbid nature, contamination is difficult to be detected...during working in the laminar airflow are also among the leading causes of contamination. These aerosols generated...cell culture process and hence eliminate one of the causes of contamination. If possible, a separate lab...bacterial and fungal contamination. Proper PPE Since humans are the major source of contamination, the prevention...

...however, the other recombinase/reporter pairs were inactive, even at very high recombinase levels. These data...multiple recombinases can be used together to designate specific patterns of recombination. Finally, Rubin...rather than complete recombination. If you’re looking to use site-specific recombination in Drosophila, read... using multiple recombinases. To make multiple, precise genome edits, the recombinases used must have ...representing the specific recombination site for each enzyme, were generated to test recombination efficiency. When...showed broad recombinase-dependent RFP expression. Even at low expression levels, recombination by KD, B2...determine the cross-reactivity of these recombinases, each recombinase and reporter construct pair were tested...

...specificity in retinal cells. To do this, the group developed a new strategy for finding retinal cell-type ...monkey, and human retinas Next, the 113 AAV promoters that were active in mouse retinas were injected into...human retinas but not in mice was low (R=0.12), suggesting that mice could be used to eliminate promoters...gene expression in retinal cell types and classes in mice, primate, and human retinas. These synthetic ...types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than using naturally occurring...they drove cell type specific expression in the retina.  Four approaches for designing synthetic promoters... with sequences upstream of the start codon of retinal cell-type specific genes from mice. Strategy #2...

...expression of other recombinases. Multiple orthogonal attB-attP pairs per recombinase may be arranged on...different recombinases are overlapping or nested on a register, then the operation of one recombinase on the... when treated with different combinations and orders of two recombinases. How might one read the state...they can control recombinase activity. For example, post-transcriptional recombinase activation via RNA...reduces the content of our paper, “Synthetic recombinase-based state machines in living cells” (1), into...explanation of how to adapt our engineered devices, recombinase-based state machines for your own experimental... about the order or timing of these factors. Recombinase-based state machines (RSMs), which take on a ...

...Having never been to China myself, I was pleased to be able to participate when the organizers of the...Dmitry Samarsky, and a very welcoming team from China’s Ribobio, the illustrious speaker list attracted...winners, one NIH Director and a co-founder of BGI-China presenting, it’s no wonder it was attended by over...were potential Addgene requestors and depositors. China is in one of the top 10 countries in number of plasmid...of Guangzhou and see some of the sites. Most fascinating to me was the outdoor market where one can shop...chemistry is complex and this work requires an interdisciplinary team.  Day two of the conference was kicked...from different species for genome engineering. The final keynote presentation was by Nobel Laureate Thomas...

...iGEM team originally used a guaB knockout E. coli strain to assay for functional decaffeination in cells...supplemented with various caffeinated beverages, but not with caffeine-free drinks. Finally, they demonstrated... pulse of blue light, and is reversible upon termination of light stimulation. CRY2clust enables faster... changes in intracellular Ca2+ levels than the original version containing CRY2 without the extra peptide..., termied soCoChR, by fusing a fragment of the kainate receptor subunit to the C-terminus of the protein...the group was able to demonstrate that while illumination of neighboring cells evokes action potentials...these APs are absent in soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish...

...plasmid-based recombinant monoclonal antibodies Resources on Addgene.org Find recombinant antibody plasmids...leading to problems with reproducibility and an inability to improve upon these critical research tools.... Addgene to create a much needed open-access recombinant antibody/affinity reagent resource for the neuroscience...and data, the platform can be expanded for any recombinant antibody tool and therefore will grow to serve... the entire scientific community.     Why recombinant antibodies? Animal-based methods for antibody...cryopreservation (Bradbury & Plückthun, 2015a, b). Recombinant antibodies are plasmid-based and offer numerous...monoclonal antibodies. You can learn more about recombinant antibodies in this blog post. Hello, NABOR! ...

...Fvasconcellos.     How are Recombinant Antibodies Made? To make recombinant antibodies, you first need..., 2015).    The Many Benefits of Recombinant Antibodies Recombinant antibodies offer numerous advantages...definition of recombinant antibodies are essential for good reproducibility. Non-recombinant antibodies ...predict a similar revolution for antibodies. Recombinant antibody technology makes it easy for scientists...components of the scientific toolbox. What are Recombinant Antibodies? Antibodies are used by scientists...scientific institutions have proposed a shift to recombinant DNA-based antibody technologies (Bradbury & Plückthun...single antibody recognizing one epitope. (Bottom) Recombinant monoclonal antibodies are encoded in plasmids...

... multiple Addgenies and most recently updated by Ina Ersing in August 2023. Resources and references ...  Ina Ersing...absence of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...Endotoxin contamination in vector preparations can alter the immunogenic properties of the final product...mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination. In the event...is performed on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed...particularly in large animal studies. Endotoxin contamination is minimized by using an endotoxin-free plasmid...

..., Diana, Dolly, Eddy, Emma, Eric, Gizela, Henry, Ina, Inna, Ishrat, Jane, Jason, Jessica, Joel, Jonathan... revenue to build reserves so the company is sustainable. The difference is what nonprofits do with their..., Lianna, Luke, Marcy, Mary, Max, Maya, Meghan, Melina, Meron, Michael Lo., Michael Le., Michel, Michelle...

...By Susanna Bachle and Ina Ersing Seeding Labs and Addgene collaborate to provide scientific resources...behind Mycobacterium tuberculosis resistance to pyrazinamide (PZA) (a drug commonly used to treat tuberculosis... different approaches, including CRISPR and recombination-mediated genetic engineering (find more information...pathogenic Klebsiella and Pseudomonas isolates causing urinary tract infections. This project aims to examine ...Adeyi at the University of Ibadan. She works with medicinal plants such as Launaea taraxacifolia (Willd), ...pathophysiological complications of diabetes mellitus. Recombinant human proteins that play a role in insulin regulation...Ernesto Valdivia Silva is a master of interesting combinations: biomedicine and astrobiology. During the last...

...plasmids at Addgene! New from the viral service By Ina Ersing We regularly add new viral aliquots from our...protease used to cleave affinity tags from purified recombinant proteins at small and large scale.  This new .... Find the plasmids at Addgene! A new dual-deaminase base editor enables concurrent adenine and cytosine...Addgene New serotypes for the Cre-dependent Flpo recombinase pEF1a-DIO-FLPo-WPRE-hGHpA, which is now also ...

...series!  This post was originally written by Didem Goz Ayturk and updated by Ina Ersing in August 2023 ... your final stocks only contain rAAV. To minimize contamination, use appropriate decontamination procedures...40% PEG solution to each 100 mL of supernatant (final concentration of 8%). If, however, you’ve collected...digital PCR (ddPCR) (Lock et al., 2014), or determination of the median tissue culture infective dose ...primers targeting the viral genome or inverted terminal repeats (ITR). You should always have a high-quality...adeno-associated virus serotype 2-derived inverted terminal repeat sequences. Human Gene Therapy Methods, ...Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors [Preprint]. Molecular...

...cloned, expressed and validated in recombinant form. The recombinant monoclonal antibodies or R-mAbs have...149451 Anti-GluK5/Grik5/KA2 kainate receptor [N279B/27R] GluK5/Grik5/KA2 kainate receptor Rat Mouse IgG2a...Anti-MMP9 matrix metalloproteinase-9 precursor [L51/82R] MMP9 matrix metalloproteinase-9 precursor Rat ...IgG2a 220411 LRRK2/Dardarin kinase, C-terminus [N241A/58R] LRRK2/Dardarin kinase, C-terminus Human Mouse ... recombinant monoclonal antibody. Cav1.2 Mouse Mouse IgG2a 225372 SynDIG2 (Mus musculus) recombinant monoclonal..., followed by the SV40 polyA tail (SV40 pA). Recombinant Monoclonal Antibody Plasmid Collection The following...IgG2a 225364 Kv2.1 pS586 (Rattus norvegicus) recombinant monoclonal antibody. Kv2.1 pS586 Rat Mouse IgG2a...

... Cre recombinase expression in striatal indirect pathway D2-MSNs Jonathan Ting Viviana Gradinaru 199776...for Cre recombinase expression in striatal direct pathway D1-MSNs Jonathan Ting Viviana Gradinaru 199775...for Cre recombinase expression in striatal medium spiny neurons Jonathan Ting Viviana Gradinaru 199774-...striatal cholinergic neurons Jonathan Ting Viviana Gradinaru 214862-PHPeB AiP14825: pAAV-AiE0441h_3xC2-minBG-iCre...striatal medium spiny neurons Jonathan Ting Viviana Gradinaru 214858-PHPeB AiP14496: pAAV-AiE0873m_3xC2-minBG-SYFP2...striatal cholinergic neurons Jonathan Ting Viviana Gradinaru 214857-PHPeB AiP14361: pAAV-AiE0452h_3xC2-minBG-jGCaMP8s-WPRE3...spiny neurons (D2 MSNs) Jonathan Ting Viviana Gradinaru 214856-PHPeB AiP14360: pAAV-AiE0779m_3xC2-minBG-jGCaMP8s-WPRE3...

...FLAG-HA-pcDNA3.1 - C-terminal Flag or N-terminal FLAG-HA for mammalian expression pNIC-CTHF - C-terminal ...vector with C-terminal mTagBFP2 tag pNIC-GST-TEV-GG - Cloning of target gene with N-terminal GST tag and...remove the stop codon for C-terminal tags and omit the start codon for N-terminal tags. And when you are designing...vectors - PCR-based C-terminal tagging in S. cerevisiae pCS2FLAG - N- or C-terminal 2xFLAG...display vector with a C-terminal Myc tag pPMW-attB - pUASp plasmid with N-terminal Myc tag and attB for ...adds an N-terminal myristoylation signal GST Protein purification pEBG - N-terminal GST for... N-terminal His6-GST-TEV for bacterial expression pDest-565 - N-terminal ...

...enhancer Viviana Gradinaru 99133-AAV1 pAAV-CAG-fDIO-mNeonGreen An AAV genome with Flp recombinase-dependent ...Cre-dependent recombination Ofer Yizhar Antibodies Addgene distributes ready-to-use recombinant antibodies... tools and emphasizes their rapid and broad dissemination to the research community. Browse plasmids ,...83894-AAV1 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent dTomato expression in forebrain GABA-ergic...83894-AAV2 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent dTomato expression in forebrain GABA-ergic...83894-AAV5 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent dTomato expression in forebrain GABA-ergic...83894-AAV9 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent dTomato expression in forebrain GABA-ergic...

...Cre-lox recombination principles, scientists have developed constructs to activate/inactivate genes when...respectively. Split Cre: Cre recombinase is split in half into N and C terminal Cre fragments (NCre and CCre...photo-activatable cre recombinase CAG AAV Bruchas 75268 pAAV-CRY2-CreN photo-activatable cre recombinase CAG AAV Bruchas...pCMV-Tag2B-NCre N-terminal split-Cre CMV Mammalian Hirrlinger 106369 pCMV-Tag3B-CCre C-terminal split-Cre CMV...pCMV-Tag2B-NCreERT2 N-terminal split-Cre-ERT2 CMV Mammalian Hirrlinger 106371 pCMV-Tag3B-ERT2CCre C-terminal split-Cre-ERT2...photo-activatable Cre recombinase CMV Mammalian Sato 122961 pcDNA3.1_PA-Cre-Y324F Catalytically inactive PA-Cre CMV...FRT: Alternative recombinase system for the control of gene expression. Flp recombinase recognizes FRT ...