... lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

...red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system is spread...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...

...individual protein has all the components necessary to: 1. Bind to a guide RNA The guide RNA, which will be ...loci. 3. Cleave target DNA resulting in a double-strand break (DSB) Cas9 and its variants have two endonuclease...positions the nuclease domains to cleave opposite strands of the target DNA. Thus, the end result of Cas9...arrays as found in the bacterial genomes are transcribed into pre-crRNAs containing both the spacer region...RNase III, the tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound...target DNA which is E) cleaved leaving a F) double-strand break. A “gRNA” is a researcher-designed hybrid...

...through the use of nanoparticles. [6].    References 1. Zhang B. 2021. “CRISPR/Cas gene therapy.” Journal...editing system to correct the cystic fibrosis transmembrane conductor receptor (CFTR) by homologous recombination...Cas9 has been used to generate mutations in a wide range of organisms, but its potential to efficiently correct...Cas-based Gene Therapy in Human Genetic Diseases.” Theranostics. https://doi.org/10.7150/thno.43360 7. Yuxuan...

... will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for...next interview or whether your receive the next grant you apply for. After all, the only value your work...high level to their team.  When you're writing a grant, this means convincing a committee that your research...collaborating with colleagues in other fields or writing grant proposals to be reviewed by interdisciplinary panels...communities gather at ComSciCon's local and specialized franchise workshops throughout the year.  Find workshops...

...into account, the digest pattern depicted in Lane 1 would be expected for a XbaI/NotI digest. The program...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...overlaps with the XbaI cut site (depicted as an orange line) because the restriction site is followed ...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...

...challenges involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the...how they started their company here. The first transgenic plant was engineered over 30 years ago, but plant... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...

... After all I was only going to be interviewed for 1 hour so having a broad enough insight was going to...promotions or further education may allow you to transition into various realms of business.  Thank you ...

...-fDIO mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg...expression, researchers often need to determine these transduction patterns empirically for their own experiments...AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon Harvey 50457 AAV2 pAAV-hSyn-DIO-EGFP Bryan Roth...system takes advantage of the extensive supply of transgenic reporter lines that supply a variety of GFP expression...

...the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two MS2, PP7, or boxB sequences...the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear processes...

...as an endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head ... knockdown efficiency across the 11 candidate transcript targets analyzed. CRISPR-Csm presents an attractive...! Colognori D, Trinidad M, Doudna JA. Precise transcript targeting by CRISPR-Csm complexes. Nat Biotechnol...

...instead conveniently provide them on your smart phone. 1. Lab Archives by LabArchives The single, most important...MSDS for a lab chemical of choice. Now, I can’t guarantee every reagent in your chemical closet is in this...this app. However, if you get quizzed randomly about a hazardous chemical and can’t find your lab’s MSDS...

...present in the data such as synapsin, hsyn synapsin-1, human synapsin, hSynap. Matching We capture multiple...categorization of our information in order to more granularly customize the search engine, and a scoping mechanism...example, matches of a name in the depositor field will rank higher than matches of the same name in the publication...

...Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One of my favorites is making the...work for almost 20 years. It is good to be a bit random because you never know which relationships will...These are all normal outcomes. If you have a broad range of mentors you will always have people you can turn...

...techniques?  Let us know in the comments! References 1. Bacteriophage P1 site-specific recombination. I. ...A double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...maintained).  Translocation: If the sites are on separate DNA molecules, a translocation event is generated... expression – placing a series of transcriptional and translational termination features with loxP sites... pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells, and/or tissue-...that there is virtually no chance of it occurring randomly. Therefore, inserting loxP sites at deliberate...Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination...

...Blood cells are not transduced efficiently with adenovirus and AAV.   Table 1: Types of viruses... Retroviruses are single-stranded RNA viruses that use reverse transcriptase to synthesize complementary...for most experimental needs, and can transduce (infect) a wide range of cell types with a high efficiency... such as a heterologous envelope or a post-transcriptional regulatory element can be used for either type...Adeno-associated virus (AAV) is a small, single-stranded DNA parvovirus. AAV can infect humans and some...response as retroviruses. This virus can also transduce dividing and non-dividing cells, making it conducive...issue. Adenovirus Adenovirus has a linear, double stranded DNA genome which is non-enveloped. It can infect...

...site. Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...the co-transduction by two viruses would lower editing efficiency, the split editors transduced at levels...capacity of 4-5 kb limits the delivery of larger transgenes, such as base editors which have a gene size ...-opsin GEVIs. With an increase in the neuron’s membrane potential, Ace absorbance increases and results...promoters, CRISPRi parts, selectable markers, transformation acceptor vectors, and sequences for genomic...plasmids using the Cyanogate Kit and subsequent transformation into cyanobacteria. Image from Vasudevan et... evaluated CreLite for cell tracing using the transgenic zebrafish model, ubi:zebrabow. In this system...

...and CasPEDIA have got you covered!     Fig. 1 – CasPEDIA Cas ID display information for a given ...Cas enzymes) cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the...cleaves ssDNA in trans. In sum, CasPEDIA documents the target substrate (DNA/RNA and strandedness) along with...Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability...features: CasID CasPEDIA will display both the cis and trans activity of each entry. For example, SpyCas9a (one...can manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.).....32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019). Evolutionary classification...

...her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization and dynamics... has recently published techniques to follow translation in living single cells. However, before CRISPR-Cas9...the Max Planck Institute for Brain Research in Frankfurt am Main, Germany. She presented a novel technology...to be invited to learn from them at the MPI in Frankfurt am Main and perform similar experiments. Visualizing...Both antibodies have a short antibody specific DNA strand added to them. Only if the proteins of interest...are in close proximity to each other can the DNA strands take part in rolling circle DNA amplification. ...microscope she likes to travel, discover new restaurants, go to concerts and bake. you can contact her...

...into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct. Image from Wen ...Wen et al., 2018). SRIRACCHA uses the Piggybac transposon to stably integrate a reporter gene that contains..., you’re ready to edit. These cells are then transfected with several different constructs for the reporter... gRNAs by simply changing the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The...plasmid.  With this set up, all successfully transfected cells will express RFP while cells containing... Lastly, once a mutant is identified, the PB transposase can easily and seamlessly remove the integrated...Knudsen C, Ásgrímsdóttir ES, Rahimi K, Gill KP, Frandsen S, Hvolbøl Buchholdt S, Chen M, Kjems J, Febbraro...