...genome-wide off-targets, since increasing PAM flexibility might also increase off-target activity. Surprisingly...gone one step further, using directed evolution to create xCas9, an enzyme recognizing a broad range of PAMs...PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited to learn more...easy, right? Yes and no. If you’re just trying to create a knockout, you might be happy targeting anywhere...precise edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10...insertion site. Base editing, another method to create point mutations, also has a strict editing window... PAM sequences and/or broader PAM flexibility. Creating xCas9 through directed evolution Hu et al. used...

...nucleobase component of the nucleoside inosine. Created with BioRender.com. Although it was a huge ...it finds a PAM, Cas9 unwinds the adjacent DNA to create a region of separate strands called an R-loop (...of the enzyme. Figure 2: CRISPR R-loop. Created with BioRender.com. Base editors take advantage...activity window restricted to the stretch of ssDNA created by Cas9 binding. Pro tip! In CRISPR, the non-target...BER process was excising the incorrect U base, creating an “abasic,” or empty, DNA site, and then replacing...in the same journal on the same day.) This team created a similar construct that they called a Glycosylase...GBE, for its reliance on the glycosylase UNG to crease abasic sites (Zhao et al., 2021). Their GBE produced...

...ones? Here, I’ll cover some screening methods to identify them. Visual screening Regardless of the RNAi-trigger...endogenous miRNA precursors can be engineered to created artificial miRNAs. The processed miRNA or siRNAs...conclusion was based upon numerous phenotype discrepancies between gene silencing and knockout methods ...RNAi in the context of a high throughput survival screen found that either technology was equivalently precise... can be divided into three fundamental steps: creation of the RNAi trigger delivering the RNAi trigger...the first stage of an RNAi-based investigation. Creation of the RNAi trigger requires a few initial considerations...these constructs include pGAPi, which allows easy creation of long hpRNA with Gateway-based cloning and straightforward...

...modular nature, the Dueber Lab created a few versions of EvolvR. To increase the mutation rate of EvolvR,...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing window of...forward genetics harnesses this at a large scale by screening diverse libraries of genetic variants. Several...nucleotides. Both the Bassik Lab and the Chang Lab have created methods that introduce diversification by targeting... integration of oligonucleotide libraries of a discrete size at the target site. The use of such libraries...occasionally introducing mutations (red nucleotide). Created with BioRender.com. The initial version of ...per generation of wild-type E. coli, while only increasing the standard mutation rate seen during DNA replication...

... Association for Creatine Deficiencies: Compensating for SLC6A8 mutations in Creatine Transporter Deficiency...SLC6A8 gene, also known as the creatine transporter. The Association for Creatine Deficiencies (ACD) is a parent-let...regeneration of ATP from ADP is creatine. In order to reach the brain, creatine needs to be transported from...Deficiency Creatine Transporter Deficiency (CTD) is a rare X-linked inherited neurometabolic condition and... and one of three Cerebral Creatine Deficiency Syndromes (CCDS) that present as global developmental delays... blood and cross the blood brain barrier using creatine transporters. However, mutations in the transporter...SLC6A8 impede this transport and result in global creatine deficiency, which is characterized by development...

...libraries here at Addgene, including cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries...determined, larger lentiviral preps can be used for screening-scale transduction of cells, resulting in cell...library. These cells can then be used for genetic screening, followed by sequencing and further data analysis...some of your plasmids didn’t amplify after your screen is completed!  Producing Your Lentiviral Library... any issues.  Keep in mind that a low MOI will increase the chances of a cell being infected by only one... one viral particle, but it also increases the chances that a cell doesn’t get infected at all. To make...as seeding density and reagent concentration.  Screening time Phew! You are now (finally!) ready to use...

...pooled libraries have been created in lentiviral systems for mammalian screens, which enables researchers... as the screen readout. This retroviral mouse CRISPR pooled library can be used for screening of cells...pooled libraries are popular tools for genetic screens, enabling genome-wide interrogation of the genes...required for cellular processes. For a successful screen, it is necessary to identify the gRNAs that elicit...are poorly transduced by lentivirus, and thus screening with available CRISPR libraries was not possible...solve this issue, Sarah Teichmann and colleagues created a retroviral CRISPR knockout library using gRNAs... from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation and activation...

...goals and the resources. Instead of creating a start up budget, create a start up list.  In this post we...award by giving you step by step instructions on creating a budget, making decisions based on that budget...budget, and negotiating in regards to the budget. Creating a budget We start by using your research specific...budgetary needs from your experimental goals. When creating your budget, you should take the following steps... that timeline Avoid writing down prices as you create your list. This helps to ensure that all of the...”, “colony”, etc. This will help save time by screening out irrelevant candidates, and, more importantly...doesn’t have to be complicated, or difficult to create this list. Start BEFORE searching for a faculty...

...gene expression. By itself, increasing the copy number of a gene increases its expression. However, if...as plasmid copy number increases, the amount of repressor protein also increases thereby intensifying repression...When they increased concentrations of IPTG, a constitutive promoter upstream of GFP increased expression...that is responsive to a repressor protein, the increase in expression due to copy number is kept in check...origins of replication, the stabilized promoters created by Segall-Shapiro et al. showed nearly no position-dependent...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a...compared to a constitutive promoter. To simulate the increase in plasmid copy number during the transition to...

... edited. Created with BioRender.com. Add silent mutations near point mutations to create 3-base (...target can increase the odds of incorrect editing outcomes, especially when MMR is inhibited. Created with ...edit. Figure 1: Prime Editing schematic. Created with BioRender.com. The pegRNA specifies both...PBS) and reverse transcriptase template (RTT). Created with BioRender.com. The first base of the ...to Cas9 with canonical sgRNA structure (right). Created with BioRender.com. What to edit The sequence... mismatched bases can often evade MMR (right). Created with BioRender.com. For specific prime editing...nicking sgRNAs ~50 bp from the prime editing nick. Created with BioRender.com. PE3b and PE5b: In this...

...engineer specific DNA edits and to perform functional screens to identify genes involved in a phenotype of interest...sequence preferences must largely be ignored. Created with BioRender.com. The hammer: Gene knockout...terminus of the protein, to maximize the chances of creating a non-functional allele. For a 1 kilobase gene...primary importance (more on this below). Created with BioRender.com. The jigsaw: Editing by ...the need for single cell cloning and subsequent screening for successful edits. This is a very time consuming...constrained by the desired location of edit; efficiency decreases dramatically when the cut site is >30nt from the...options that can greatly expand gRNA options. Created with BioRender.com. Two newer technologies ...

...activation. Created with BioRender.com. Tet1 demethylase Ronggui Hu’s lab has created pdCas9-Tet1-...targeted demethylation is associated with increased transcription. Created with BioRender.com. Tet1 initiates...the catalytic domain of p300 acetyltransferase increases levels of H3K27ac histone modification at specified...This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation and activate transcription. A lentiviral...DNA oxidation and repair factors that allow for increased gene activation at the targeted site compared ... of transduced cells. Grant Challen’s lab also created constitutive (pCMV-dCas9-D3A) and Tet-dependent...is associated with transcriptional repression. Created with BioRender.com. DNA Methyltransferase MQ1...

...closer to an innovative resolution. Creative negotiation can also decrease or entirely eliminate the feeling...and everyone leaves unhappy.   Negotiation as a creative problem-solving process Although at first glance...negotiating, what if we instead approached it as a creative problem-solving exercise? Asking questions like...involved and be used as a guide to point you toward a creative solution where everyone wins. What are the underlying...but still be beneficial to both.  Coming up with creative solutions where everyone wins can initially feel...person. It can especially be difficult to think creatively when you are in the middle of a frustrating situation...practice will make negotiating feel more natural. Creative negotiation skills will help you strengthen relationships...

...of themselves. This increases the number of copies of the TE while also increasing the size of their host...mutagenesis screens that detect loss-of-function or gain-of-function mutations. In these screens, the transposons...transposon for a mutagenesis screen. For example, piggyBac and Tol2 are best for screening promoters and enhancers...Then they can be inserted into new target sites. Created in BioRender.com.   Class II TEs: DNA transposons...which produces a target site duplication (TSD). Created in BioRender.com.   Autonomous vs. non-autonomous...capacity >100 kB, although integration efficiency decreases with cargo size. Sleeping Beauty has a close-to-random...can deliver 10-11 kB to mammalian cells without decreased efficiency, with a maximum cargo capacity of ~...

...during your in vivo work is to screen you gRNA in vitro first. In vitro screening is a simple and fast method...Researchers are now using this system in ever more creative ways, (Wang et al., 2013, Cho et al., 2014). There...has been quite successfully adapted for use in screening experiments as well). Therefore, if you’ve started...previously validated gRNAs at Addgene. In vitro screening for the best gRNA Figure 1: Validation ...gRNAs to Target Mouse Genes To begin the in vitro screening process, you should first develop primers to the...same time, and choose the most effective ones to increase your chances of gene modification in vivo. Find...embryos to develop. Once they are born, it’s time to screen them for your desired edit. Good news! Remember...

...compose the secretory pathway, endocytic vesicles, mitochondria, lysosomes or they are secreted into the ...your cloning strategy and decide whether you will create an N- or C-terminal fusion based on your experimental...residues form inappropriate disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs ...and a sequence intended to localize them to the secretory pathway, a dark pool that is mislocalized to the...misfolded, blocked from proceeding through the secretory pathway, and retained in the ER. FPs have been...recent publication illustrated the quantitative increase in fluorescent signal achieved when utilizing ...optimized moxFP as opposed to standard FPs. The increase in signal is not due to a brighter FP, but rather...

...the process that was used to create this important collection.  Creating the Plasmids To begin constructing...were able to greatly increase the number of functional constructs that were created. This greatly limited...construction and validation of some of these tools created by the Trimmer Lab that will be the first antibodies...that had already been thoroughly validated. The creativity in employing a variety of cloning techniques ...from cryopreserved hybridomas, immortalized cells created by fusing an activated B cell that produces the...VH) chain sequences. Fusion PCR was performed to create an amplicon that  was cloned into a plasmid backbone...plasmid to express the target antigen could be screened in a high-throughput manner. Cells were labeled...

...you can be incredibly frustrating. Clarifying the words —> decreasing frustration —> increasing the happiness...in meetings and 2) creating a pictorial guide for our engineers (Fig. 2). Creating this guide was relatively... an engineer that you want the LIMS software to create a barcode for your plate, you will get in trouble...happening. Our solution for this problem was the creation of an Addgene lexicon. The base lexicon was and...received formal definitions in our lexicon. We created documents and presentations, gave real life examples... field or technology in a consistent way. Just creating a lexicon was not enough. We also had to enforce...the terminology of a new field. Annotate: When creating important documentation, including SOPs or project-related...

...about pooled library screening, read our recent blog post on Genome-wide Screening Using CRISPR/Cas9. If...metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that...interest to mammalian cells. It was designed to have increased lentiviral titer as compared to previous lentiviral... population of cells as part of pooled library screens (1,2). The human and mouse GeCKO libraries, targeting...lentiCRISPR v2 is key to its efficient use in screening experiments; large amounts of virus are required...you get full library coverage in pooled library screens. Availability and Use LentiCRISPR v2 is available...Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening in human cells." Science 343.6166 (2014): 84-87...

...NG-Cas9 (PAM: NG) to create NG-ABE8.xm/d and the S. aureus Cas9 (PAM: NNGRRT) to create Sa-ABE8.x-m/d. With....xm/d showed a 1.6x median increase and Sa-ABE8.x-m/d a 2x median increase in editing frequency over ABE7.10...combination with a synthetic library of TadA sequences to create the eighth generation of adenine base editors, ...selection. As a result, 8 mutations in TadA could create all three edits and were thus more efficient. Informed...behavior to ABE8.x-d. Overall, ABE8s showed a median increase of 1.94x editing activity compared to ABE7.10....ABE8.17-m/d, ABE8.20-m/d) to evaluate further.  The creation of accidental insertions and deletions (indel)...ABE7.10 can be a problem. Therefore, the authors created ABE8 constructs using a catalytically “dead” S....

...where DNA is being created, such as PCR amplification or bacterial growth. Created with BioRender.com....effort into our quality control (QC) processes to screen for and reduce these errors. These processes allow...  Figure 2. Addgene’s basic QC process. Created with BioRender.com. Once Addgene receives deposited...process ensures that any potential sequence discrepancies can be caught prior to distribution. Catching...issue? At Addgene, a QC issue is defined as a discrepancy that we believe may affect the function of the...decisions when analyzing the wide variety of discrepancies encountered in plasmids deposited to Addgene...resolve. Sequencing and assembly issues Sequence discrepancies are not always a result of mutations in the ...

... type of primer pair is also great for screening clones created with the same backbone but that contain...it's in the plasmid Can be used to screen clones that were created with the same backbone because primers...Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally...is, allowing you to use the same primer pair to screen for the presence of many different inserts. The...one primer anneals to the insert. A simple way to create this type of primer pair is to mix-and-match insert-specific...test your colony PCR primers before using them to screen colonies. The best way to do this is by using your... background, the more colonies you will need to screen.   Saving clones for later culture: At this point...

...ve learned that increased accessibility is achieved through interconnected incremental changes. It was...When Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids,...identified and offered a number of solutions that increased accessibility, such as using couriers that refill...In some cases, the distributors broadly helped increase access in the country, and we can see a growth...re continuing to use our data and resources to increase accessibility to researchers around the globe....the materials in our repository. We started to increase accessibility to our repository by making it easier...Bacterial Transformation How to Design a Primer Creating Bacterial Glycerol Stocks Inoculating a Liquid...

...article.) Modifying AraC to create BLADE The team modified AraC to create BLADE by replacing its dimerization...images created by the team (Fig. 2). These images are based on the concept of bacteriographs created about...bacteriograph of Michelangelo's "The Creation of Adam." This bacteriograph was created using 160 individual images...dimerize and activates transcription. The team created two constructs, among others: pBLADE-mCherry:...about a decade ago. To create these images, the scientists used a strain transformed with pBLADE expressing...instead of mCherry under the PBAD promoter. They created a bacterial lawn on an agar plate, put a mask over... that Di Ventura’s lab and Khammash's lab have created and shared with Addgene. You can find all the plasmids...

...Cancer cells find a way to grow haywire, access and create more vasculature to feed themselves, use the blood...the tumor microenvironment - a vital atmosphere created by the cancer cells to shield themselves from attack...cells of epithelial origin. Suppressing function, creating tolerance, and evading recognition are a few methods...cells into regulatory cells to suppress function. Creating Tolerance T-regs in the tumor microenvironment...HIF-1), which promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates...signals released from the microenvironment help create tolerance beyond direct proximity to the tumor;...cells (DCs) and macrophages (M1 and M2) that help create welcoming environments for tumor cells spreading...

...mutations increased the accuracy of editing by up to 10-fold per allele. This is a 100-fold increase in editing...incorporate homo- or heterozygous mutations. 1. Increasing HDR accuracy with CRISPR/Cas-blocking mutations...find the 0.2-0.5% correctly edited alleles, when screening reasonable amounts of single cell clones. Furthermore...tedious business of iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000... together with the intended sequence change and screen a few hundred clones to find the right one Step... few hundred. From these two steps, you’ll be screening a total of a few hundred clones - Still better...efficiency of mutation incorporation falls with increasing distance from the cut site. If the distance is...

...recombination, increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability...that make major improvements such as those that increase plasmid yield and/or DNA quality. Table 1 below...sensitive. dnaJ Mutation in a chaperonin gene Increases the stability of certain expressed proteins ...methylate DNA lac Lac operon mutations Blue/white screening of clones   lacIq lac repressor overproduced...derivative. General cloning and storage, blue/white screening, leucine auxotroph. F- endA1 recA1 galE15 galK16...cloning and storage of common plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR...General cloning and plasmid maintenance, blue/white screening, partly restriction-deficient; good strain for...

...encoding Gag and Pol, increasing the number of recombination events necessary to create a replication-competent...Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...shows how the lentiviral genome was condensed to create the 2nd-generation lentiviral system (Figure 2)...safer than the original system, the possibility of creating a replication-competent virus via recombination...possibility and enhance biosafety, Dull et al. created the 3rd generation system (Figure 3), which differs...Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the lab - the Trono lab’s lentiviral...both for targeted modification and genome-wide screening.   Lentiviral vectors in the clinic Since lentiviral...

...Marker B. Created with biorender.com.   The goal of flow cytometry is to use increasingly more stringent...populations that are B+ and one population that is B-. Created with biorender.com.   If it’s hard to conceptualize...imagining (or drawing) a quadrant over the dot plot. Created with biorender.com.   Of course, this is quite...al., 2017 to identify mDCs and pDCs. Used under Creative Commons license.    In the third plot, anti-CD14...visualize the different populations. Used under Creative Commons license.    Pro tip! Subtypes of immune...the percentage of cells in each gate. Used under Creative Commons license.    Contour plots  FACS data...data from Jhunjhunwala et al., 2015. Used under Creative Commons license.  Histograms  For analysis...

...CBEs). But unlike CBEs, CGBEs then excise the U to create an abasic (empty) DNA site using either fused or...nucleobase component of the nucleoside inosine. Created with BioRender.com. Inosine excision leads ... able to edit A → Y (Y = T or C) in 67.17% of screening cells (Tong, Wang, et al., 2023). Based on this... for A → X base editor (X = any nucleotide). To create a more specific A → C base editor (ACBE), they ...Black arrows indicate natural cellular processes. Created with BioRender.com. The first deaminase-free...either to directly accomplish the base edit or to create the substrate for a base excision repair pathway...nCas9. First, MPG removes the G base from DNA to create an abasic site. Like in most other base editor ...

...in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes. For... storm. While Cas9 has been most widely used to create specific edits in DNA, there has also been significant...inducible upon environmental triggers. This is done by creating a split dCas9 protein that can reassemble when...the best uses for Cas9 activators is in genetic screening. gRNAs targeting every gene in the human genome...interactions. Additionally, large scale CRISPR-based screens can be used to gain insight into protein structure...focused advancing scientific discovery through the creation of innovative technologies. To date his group ...of cancer therapeutic targets using CRISPR–Cas9 screens. Nature 568:511–516 . https://doi.org/10.1038/s41586...

...nuclease domains, researchers created the CRISPR nickase. Nickases create a single-strand rather than a...generated by the nickases; D10A creates 5’ overhangs and H840A creates 3’ overhangs in a PAM-out design...Conversely, the H840A mutation in the HNH domain creates a non-target strand-cleaving nickase. Instead of...strands bluntly with WT Cas9 and one gRNA, you can create a staggered cut using a Cas9 nickase and two gRNAs...other? The gRNAs must target different strands to create a DSB, but this can be accomplished with either...an individual gRNA with WT Cas9, repair levels decrease rapidly 10 bp from the cut site. So if you can... you can’t obtain high HDR efficiency. Nickases create a staggered cut - could this system mediate repair...

...autophagosomes with lysosomes leads to an increase in yellow puncta but a decrease in red puncta. Find the mRFP-...RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence suggests an increase in autophagic...flux, while an increase in GFP fluorescence relative to RFP fluorescence suggests a decrease in autophagic...yeast or the lysosome in mammalian cells. This decrease in pH quenches SEP but not DsRed, leading to emission...Therefore, the formation of autophagosomes leads to an increase of yellow puncta (GFF positive/RFP positive), ...fusion of an autophagosome with a lysosome. An increase in autophagy leads to more yellow and red puncta...

... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...serine with dispersed hydrophilic residues for increased solubility (Monnier et al., 2013). The linker ...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...applications where longer retention times often increase therapeutic efficacy (Ahmed et al., 2012). How...and VH sequences. Phage display also allows for screening scFv generated from the B cell antibody genes ...the ways antibodies are. scFvs are even used to screen for new antibodies. To do this, antibody variable...variable domain sequences are expressed as scFvs and screened with in vitro assays to select for strong binders...

... are isolated from the resulting colonies, and screened for the desired modification. Finally, the positive...final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of interest...multimerization (purple) which is evident by an increase in the size of the lower band (portion of the ...mutation). Similar results can be obtained with PCR screening using the primers specified by the arrowheads ... vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward and reverse...Template A high purity plasmid prep significantly increases the success rate of site directed mutagenesis....secondary structures of the DNA template, and may also decrease the annealing temp of the primers. Because you...

...14 and 21 to create GEmCherry2. The Denham lab made several other small changes creating 2 other versions...Cas9 activity assays. Recently scientists have created CRISPR fluorescent reporter assays to quantify ...CRISPR/Cas-stimulated HDR activity) The Madison lab created SRIRACCHA in 2017 to enrich and identify desired...H2B-GFP. This visual read out allows for easy screening of different gRNAs by simply changing the gRNA...plasmid that contains Cas9. The Madison lab also created an inducible SRIRACCHA system (iSRIRACCHA) where... H2B-GFP expressing cells exhibited a 2-4 fold increase in the number of endogenous indel edits when compared... sites and activity (Knudsen et al., 2018).  To create GEmCherry2, the Denham lab first incorporated an...

... the injunction. While much of this decrease is due to an increase in the number of universities’ whose...postdocs who were expecting a salary increase have had that increase cancelled. Rounding out the remainder...payment for working more than 40 hours per week increasing from $23,660 to $47,476 per year. This post discusses... of postdocs who weren’t going to get a salary increase prior to the injunction still won’t after the ...issues such as childcare using money from their increased salaries. In addition, salary uncertainty may ... costs of living would get much better salary increases that those in less densely populated areas. This...Gary's Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating for Science...

... of interest. Although efficiency may decrease with an increased number of fragments, or the ligation ... cleave outside of their recognition sequence, creating four base flanking overhangs. Since these overhangs... fragments, these problems can be overcome by screening a higher number of potential clones. Golden Gate...the ordered assembly of multiple DNA fragments to create TAL effector nucleases. These plasmids were designed...This cloning strategy not only makes it easy to create a single gRNA-expressing plasmid, but it can also...fragment: PCR-based amplification can be used to create silent point mutations at internal recognition ...Whether you are using the Golden Gate method to create CRISPR/Cas9 constructs, assemble standard plasmids...

...challenging for amateur scientists. Planarians, wormy creatures commonly found in freshwater ponds, provide a ...promoters to allow for bidirectional transcription to create dsRNA. Students can produce DNA templates via PCR... PCR off of the same plasmid and use these to create riboprobes for in situ hybridization experiments.... as how to perform worm amputations. They also created an online companion resource called “cutting class...within an embryo or tissue. Once the riboprobes are created, the WISH protocol involves preparation of the ...as a result, these cells turn blue. Students can create synthetic riboprobes off of linearized DNA templates...from their organism of interest, use the RNA to create a complementary DNA (cDNA), and then use PCR and...

...protein in your hand? At 3D Molecular Designs, we create physical models of proteins using 3D printing technology... real”.  We sometimes forget that the models we create are around 30 million times larger than the real...were soon developed that allowed researchers to create stunning, interactive images of their proteins....physical models of protein structure? We have been creating physical models of proteins for researchers for...educators and students use molecular models?   We created the Center for BioMolecular Modeling (CBM) in 1999... 1999 with the expressed purpose of creating physical models of proteins and other molecular structures...researchers to learn about their research while creating physical models of proteins that are being investigated...