... Carrier Proteins (SLCs) from the RESOLUTE collection AAV Packaging from Addgene Parkinson's Research ...SARS-COV-2, ACE2, and CRISPR tools Fluorescent Proteins from the Michael Davidson collection Protein Labeling...available to industry scientists. ID Plasmid Gene/Insert PI Kits Kit name Type PI Description High Complexity...Screening Messer and MJFF Created for research in Parkinson's Disease; For target validation and drug design...agreement between the recipient and the depositing institution(s). It governs the transfer of biological materials...distribute plasmids on behalf of the depositing institutions. These agreements indicate the language which... by an authorized signatory at the recipient institution. An authorized signatory is someone who can execute...

...is fused to the DNA backbone that contains a target sequence against the fluorescent reporter in an inverted...single hairpin dsRNA is produced that contains sequence against your gene(s) of interest and the reporter... and Biotechnology from Worcester Polytechnic Institute. What is RNAi? The loss-of-function (LOF) experiment...downregulation, or “silencing,” of any gene that contains sequence complementary to the dsRNA (Wilson and... is facilitated by DNA constructs that enable insertion of ~400 bp complementary to your gene target as...whereas shRNAs are exogenously derived. Importantly, instead of the assortment of siRNAs generated from a long...outside of limited, single cell protoplast analysis. Instead, microscopic plants must be visually identified...

... enzymes - Cas13a contains two HEPN domains, whereas Cas9 uses HNH and RuvC domains to cleave target DNA... HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a molecule results in...SpCas9 is not the only game in town, and other Cas proteins like SaCas9 and Cpf1 can circumvent the limitations..., and the various applications of Cas13a. The origins of Cas13a: An RNA cleaving CRISPR nuclease Cas13a...form of “bait” to identify new CRISPR-associated proteins within the bacterial genome. From this analysis... DNA. The HEPN domains within Cas13a are essential for RNA cleavage, consistent with known roles for HEPN...Comparison on Common CRISPR Enzymes Name  Enzymatic Domains Guide RNA Target PAM (PFS) Cleavage Mechanism ...

...nickase. The fusion enzyme is capable of installing targeted insertions, deletions, and point mutations using...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter Selectable...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Drosophila ID Plasmid Gene/Insert Promoter ...edited 3’ flap is excised, the target sequence remains unchanged and available as a substrate for another...small prime editors with optimized RT and/or Cas9 domains PE7 - addition of RNA-binding exonuclease protection...enzyme, selectable marker, and whether the plasmid contains Cas9. ID Plasmid Expression System Promoter Guide...

...Includes many N- and C-terminal fluorescent proteins Insect Sutherland Lab Plasmids - For multicistronic...different fluorescent tags for you to create fusion proteins with your gene of interest. Special... Monomer not a Monomer? Background Fluorescent proteins are genetically encoded tools that are used extensively...have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set of colors. Addgene...different fluorescent tags for you to create fusion proteins with your gene of interest. These are organized... at high concentration, so this is noted here. Proteins that are derived from GFP and contain the A206K...information on the properties of many of these proteins, see: Cranfill et al. , Nature Methods 2016 Jul...

...users can submit new fluorescent proteins, add data to existing proteins when missing, or edit inaccuracies... specific protein (for instance: EGFP).  Users can “favorite” certain proteins, or create custom protein...recognized by the 2008 Nobel Prize, fluorescent proteins (FPs) have become one of the most indispensable... one might require when making such a decision remains spread across literature in publications that introduce...community resource for information about fluorescent proteins.  It was designed with the goal of encouraging...Protein Properties provides a master list of all proteins in the database, and the Interactive Fluorescent...moderate your own version of fpvis.org with the proteins of your choosing!), and they can be downloaded...

...more fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be diluted out...excited to see the many creative uses of fluorescent proteins. We’re happy to announce the three winners of ...by fluorescence-activated cell sorting into four bins based on their eGFP:mCherry ratios. Genomic DNA ...response to warfarin. “These novel methods will be instrumental in interpreting human variation at large scales...making personalized medicine a reality,” Chiasson explains. Elizabeth Unger: Developing a serotonin sensor... sensor. However, there are no known bacterial proteins that bind serotonin. Unger redesigned the binding...patient and rapidly test the variants in vitro against different drugs and dosages.  Michal Bernach: Developing...

...candidates for further development. They inserted an octet of MS2 hairpins into the gRNA tetraloop and went through...-head against CRISPRainbow. Both tools were targeted to the FBN3 intronic repeat which contains 22 copies...the genomic loci of interest. Pairs of identical hairpins within one gRNA will result in a blue, green, ...red readout. But gRNAs containing two different hairpins will produce a secondary color (yellow, cyan, ...hairpin combination. A gRNA containing all three hairpins produces a white color. While CRISPRainbow allows...CRISPRainbow labeling. Two MS2, PP7, or boxB sequences inserted into the sgRNA generates blue, green, or red colors...induce its fluorescence. They found that aptamer insertion in the gRNA tetraloop (a four base hairpin loop...

...recombinant proteins a cinch. This is because it doesn’t secrete its own endogenous proteins, making the...affordable, and it grows fast to boot. The yeast strains S. cerevisiae and S. pombe have dominated the research... between two of the most frequently used yeast strains: S. pombe (fission) and S. cerevisiae (budding)...established in yeast, including a widely used Parkinson’s experimental model. In fact, approximately 25%...differences between K. phaffii and other yeast strains.     S. cerevisiae S. pombe K. phaffii...and bears more similarity to the ancient yeast strains and metazoan cells.   Applications for K. phaffii...was co-opted for the expression of recombinant proteins by the pharmaceutical and biotech industries as...

...ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids... vectors for tag insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR plasmids ...includes CRISPR, RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids. Large collections...briggsae , etc.) gene or sequence: ID Plasmid Gene/Insert PI Don’t see your gene listed? Know of some great...elegans genome engineering - Bob Goldstein Lab. Contains a set of plasmids for building homologous repair...Transgenesis Kit - Erik Jorgensen Lab. This kit contains 29 plasmids used to generate transgenic nematodes...Caenorhabditis Genetics Center (CGC) - The CGC collects, maintains, and distributes stocks of C. elegans. Silencing...

...majority of Type VI-D loci contain accessory proteins with WYL domains (named for the three conserved amino acids...domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins an intriguing... near a putative effector nuclease. The Cas13d proteins they identified have little sequence similarity...Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the Cas13 superfamily. Yan et...advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are...intriguing counterpart to anti-CRISPR proteins that negatively modulate the activity of Cas enzymes, some...specific editor in mammalian cells Not all Cas13d proteins are functional in mammalian cells, but Konermann...

...podcast segment The human proteome contains a breadth of modular domains that facilitate protein-protein ...for RNAi. The Vellai lab’s new plasmid, T444T, contains T7 terminators adjacent to the T7 promoters to...TurboID and miniTurbo Article contributed by Eric Perkins  Listen to the TurboID and miniTurbo podcast segment... study of the distribution and interactions of proteins within these structures. Enzyme-catalyzed proximity...distribution and interaction networks of specific proteins in living cells. Until recently, the enzymes most...APEX2 and BioID. APEX2 is fast, tagging proximal proteins within minutes, but requires H2O2, which can be... tailored for the detection of other norovirus strains and even other viruses by modifying the sequence...

...possible to generate antibodies against many types of molecules including proteins, lipids, and carbohydrates... post we’ll focus on antibodies that recognize proteins. Figure 1: Steps in the generation of hybridomas...hybridomas come from animals that were immunized against the target protein. Animals are first injected ...special medium called HAT. This medium selects against unfused myeloma cells and antibody-producing B ... will exhibit off-target binding to unintended proteins. This is because monoclonal antibodies recognize...amino acid sequence will be shared across other proteins. Conversely, polyclonal antibodies - which are...recognize epitopes that are also present in other proteins. Because of the specificity of monoclonal antibodies...

... This means recombinant proteins can aggregate or precipitate out as insoluble granules. We overcame this...decidedly uncomfortable 45°C). Their more adventurous cousins are the extremophiles - the legends who not only...of processes. The food processing industry, for instance, employs psychrophilic enzymes for milk processing...enzymes for the host The expression of recombinant proteins for industrial or research purposes requires stable... standard conditions. However, one of the many proteins which are not compatible with regular mesophilic...temperatures suitable for the expression of psychrophilic proteins. The primary problem we wanted to overcome was...folding and expression of recombinant psychrophilic proteins. As the figure below shows, E. coli expressing...

...available here for free!) contains strategies, hints, anecdotes, and insights. After I wrote the first ... at the National Institutes of Health, was an Associate Professor at Albert Einstein College of Medicine...Hughes Medical Institute. His interests include the quality control of secretory proteins in the Endoplasmic... and mentoring to several people that found my insights helpful. All went on to get faculty positions ...specific questions and experiments. Nobody EVER complains that a seminar was too easy to understand. Make... Distinguishing correctly folded and misfolded proteins is essential for human health. Failure to do so...terms. "Implications for human disease" is vague. Instead, consider "A mechanistic understanding of this ...

... HIV so that infected cells will produce viral proteins and die, either through cytotoxicity or the immune...transcription, but with two additional activation domains (MS2 and p65) to enhance transcriptional activation...expression. In cell lines that can produce HIV-1 toxic proteins, CRISPR/Cas9 SAM caused apoptosis, indicating ...cell and 2. prevent reinfection of those cells. Kaminski et al. designed gRNAs to target the HIV-1 5’ and...Importantly, no off-target effects were detected. Kaminski et al. observed no negative effects on cell viability...up to 7 mismatches to the gRNA target sequence. Kaminski et al. next tested their procedure in HIV-1 infected...expanded, and infected with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly ...

...endogenous proteins is an exciting strategy, it is difficult to apply this technique to oncoproteins that lack..., a postdoctoral fellow at Dana-Farber Cancer Institute. Targeted protein degradation In the Bradner and...synthesize compounds that enable selective removal of proteins-of-interest from the proteome. Rather than inhibiting...degraders” recruit the proteasome to destroy targeted proteins. We previously developed small molecule degraders...that achieve selective degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and...linking small molecules that bind these target proteins to other small molecules that bind an E3 ligase... molecule) and expression of FKBP12F36V-tagged proteins. The dTAG molecule induces ternary complex formation...

...composed of four peptide chains (two identical heavy chains and two identical light chains) each with a constant... The chains are covalently linked to one another by disulfide bonds between the two heavy chains and between...are composed of just the variable domains of the heavy and light chains of an antibody. Arguably not a “...chain and one light chain. When assembled, the four chains make a Y-shaped protein that can be divided into...portions of the constant regions of the two heavy chains. In the lab, this is the region that determines...fragments are produced by proteolytic digestion that retains an antibody’s disulfide bonds within the hinge ...the fragment could bind not only the endogenous proteins you are trying to block but your primary antibody...

...these three kinases, photoswitchable Dronpa domains were inserted at sites homologous to the locations they... other kinases and proteins. Zhou et al suggest that more photoswitchable proteins could potentially be... be created by inserting pdDronpa domains into loops of a protein of interest. If you plan to try this...ninja term for vanishing, and pa= photoactivation) domains on either side of the active site of a constitutively... Learn More about Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox Control...reporter plasmids Find photoactivatable fluorescent proteins in empty backbones in the Michael Davidson Collection...Photoswitchable Fluorescent Proteins ...

...transversion editors. New DNA repair proteins fused to CGBEs Instead of using UNG, the Wei Leong Chew Lab...repair proteins fused to CGBEs The David Liu lab also experimented with adding different proteins to the...approach to improving C → G editing. Instead of fusing DNA repair proteins to the base editor, they added the...inhibit BER during cytosine-to-uracil editing, and instead introduced its opposite, a uracil DNA N-glycosylase...excising the freshly edited U bases — but E. coli instead filled those abasic sites with A's, producing C...Mycobacterium smegmatis, termed UdgX, they found three proteins whose fusion improved the CGBE: DNA polymerase...(RBMX) (Koblan et al., 2021). Several of these proteins had additive effects when multiple were included...