Telepathy and Mind Control: From Science Fiction Movies to Reality?

Posted on by Terri Sundquist

BrainInterfaceThere is something very futuristic, and perhaps scary, about the idea of nonverbally transferring one person’s thoughts to another person, especially for the purpose of controlling or influencing a person’s actions and behaviors. Maybe that’s why telepathy and mind control are favorite topics of many science fiction movies. However, there are times when direct, nonverbal transfer of thoughts would be advantageous, for example when communicating complex concepts or feelings that are difficult to convey. Direct transfer also would circumvent the need to translate information from one language to another. For these reasons, scientists are currently developing technologies to allow such thought transfers. A recent PLOS ONE article describes a simple brain-to-brain interface in humans and shows how this interface can be used to capture a thought generated by one person and communicate that information directly to the brain of a second person and elicit a physical response (1).

Continue reading “Telepathy and Mind Control: From Science Fiction Movies to Reality?”

6 + 1 Ways Dual-Reporter Assays Can Save Your Data

Posted on by Kyle Hooper

Updated 02/12/2021

Dual-Reporter-Assay

Transient transfection is often used to perform reporter assays. We have advocated using a dual-reporter system for decades to normalize the data obtained and gain a clearer understanding of your results.  The experimental reporter should vary with treatment and the control reporter should vary little with treatment. The control reporter thus serves as a marker to help you understand the relative activity of your experimental reporter. The bioluminescent Dual-Luciferase® method allows for sequential detection of the second reporter in a single sample providing a simple two-step normalization method. Here are seven ways in which dual-reporter assays help you avoid misinterpreting results.

Simply comparing the ratio of the experimental to the control reporter can resolve differences in:

  1. Number of Cells/Well: When manually pipeting cells into a 96-well plate, there is always a chance of having variable numbers of cells in each well. This variation is cell number will affect the experimental and control reporters equally, so the ratio of experimental:control reporter activity will eliminate false interpretation of the experimental data–whether it affects an entire row or column on the plate or individual wells.
  2. Transfection Efficiency: The variations in transfection efficiency will equally affect both the experimental and control reporters so the ratio of activity in dual-reporter assays will normalize the data.
  3. Cell Viability: Often, reporter assays look at the dose response curve of a particular compound with regard to gene expression. Ideally, if a compound causes a change in the experimental reporter the control reporter will demonstrate little effect. However, if the compound is toxic, both the experimental and control will be altered and the ratio will tell you whether the compound truly affects reporter activity or just kills the cells.
  4. Lysis Efficiency: When lysing a plate of cells, you could encounter situations where rows or columns lyse differently, especially if you are using manual disruption or get interrupted mid-plate. The difference is lysis will affect the experimental and control equally so the ratio will remove the variation.
  5. Temperature: Ideally, a plate should be equilibrated to ambient room temperature before proceeding to the reporter assay. Plates can cool at different rates or researchers anxious to record data may read the data early. Temperature variations will affect both reporters so the ratio will limit the affect on the data.
  6. Measurement Time: Repetition of data is a hallmark of good science. You are often called upon to repeat experiments sometimes days or weeks apart. Let’s say you repeat your experiment one week after the initial experiment. The first time you measured the response, you waited 10 minutes after reagent addition to read, this week you waited 30 minutes. This will affect both reporters equally and therefore the ratio will allow you to more easily compare the data from this week and last week.

Bonus Benefit from Dual-Luciferase®, Dual-Glo® and the NanoGlo® Dual Luciferase Reporter Systems: No Lysate Splitting: Promega dual-reporter assays are designed for same-well multiplexing so there is no chance of variations creeping into your data due to unequal splitting of the cellular lysate to measure two separate reporter activities.

Since the introduction of the first bioluminescent dual-luciferase assay in 1995, this approach has been used in countless studies to advance our scientific understanding of cellular gene regulation.

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Biotechnology Youth Apprenticeship Program Fosters Young Scientists

Posted on by Barbara Bielec

Student working in laboratory.
Photo credit: BTC Institute.
Ellyn Lepinski is an intern at Promega who started her biotechnology career path five years ago as a high school junior taking a course from the BTC Institute (www.btci.org) as part of the Biotechnology Youth Apprenticeship Program.

Ellyn credits the program with helping her achieve her goals:

“Over the course of two years in which I was a Youth Apprentice, I obtained numerous skills, both inside and outside of the lab. I gained valuable scientific experience, including techniques like gel electrophoresis, nucleic acid purification, PCR, SDS-PAGE, Western blotting, cell culture and more.

On a personal level, I became very close with other students in the class and with our instructors, Barbara Bielec and Chad Zimprich. Everyone involved was always very approachable and willing to help with both laboratory tasks and in terms of giving advice for the future.

Through the program, I was placed in Dr. Que Lan’s entomology lab at UW-Madison, beginning in 2009. While there, I worked on a project involving sterol carrier protein-2, a protein involved in cholesterol uptake in mosquitoes.Notably, I am still working in Dr. Lan’s lab, however my research focus has shifted to bacterial fermentation. In between working in Dr. Lan’s lab, I also worked at the Forest Products Laboratory (USDA).

Additionally, this past June, I began an internship at Promega in the Scientific Applications department. Here I work to develop new applications for existing projects. This November marks five years of laboratory research for me, which would not have been possible without the Youth Apprenticeship Program and everyone involved. In addition to the specific labs that I have had the opportunity to work in, my experience in the Youth Apprenticeship Program has allowed me to emerge as a leader in my college lab courses. The program has clearly made a phenomenal impact on my life and is something I am very grateful for.”

Photo credit: BTC Institute.
Photo credit: BTC Institute.
Since 1993, the BTC Institute in partnership with the Dane County School Consortium has helped make such opportunities possible to nearly 300 students from public schools throughout Dane County. The program includes a paid apprenticeship in an industry or UW-Madison research lab and specialized instruction. In addition to being paid for their work, students receive high school credit for their participation in the worksite and the specialized biotechnology course held at the BTC Institute.

One aspect of the program that makes it so effective and unique is the amount of time that students spend working. Youth apprentices who start as juniors in the program must work 900 paid work hours to earn the Science, Technology, Engineering and Math (STEM) Skill Standards Certificate from the State of Wisconsin, youth apprentices who start work as seniors must earn 450 work hours. Students have had employment at a variety of companies and UW-Madison research labs, a few examples that have hired multiple apprentices include Genus PIC (ABS), MOFA Global, Promega and laboratories in the UW-Madison Departments of Bacteriology, Biochemistry, Entomology, Genetics, Horticulture, Plant Pathology and Surgery. Many of the students, like Ellyn, continue to be employed by their worksite long after they graduate from high school—proof of how effective this program is in helping to create the next generation STEM workforce.

Each year the BTC Institute hosts a Youth Apprenticeship Program preview night for all of the Dane County youth apprenticeship options: biotechnology, automotive technician, health services, and many more (www.dcsc.org). This year the preview nights will be held February 24 and 25 starting at 5:00pm. Students in grades 10 and 11 who are interested in learning more about the program are encouraged to attend one of the evening sessions with a parent.

Genome Editing and Reporter Technologies Enable Endogenous Pathway High-Throughput Assays

Posted on by Promega

ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;
ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;


This article review was written by guest author, Amy Landreman, in the Cellular Analysis and Proteomics Group at Promega.

Charcot-Marie Tooth (CMT) disease is one of the most common inherited neurological disorders affecting approximately 2.8 million people worldwide. The most common form of CMT, CMT Type 1A, is caused by a 1.5Mb genomic duplication on Chr17 that results in trisomy of the critical myelin gene Peripheral Myelin Protein 22 (Pmp22). The extra copy of Pmp22 results in excessive PMP22 protein causing the neurophathy associated with CMT type 1A. Although there is no way to remove the extra copy of the gene, even subtle decreases in Pmp22 expression have shown promise against this inherited neuropathy in laboratory models.

In a recent paper, Inglese et al. 2014, describe an interesting new approach used to identify compounds that effectively decrease Pmp22 expression using a novel gene editing strategy and reporter-based screen. Their challenge was to create an assay that accurately represented endogenous Pmp22 expression including both transcriptional and post-transcriptional regulatory mechanisms, while maintaining the sensitivity required to detect subtle changes in expression in a loss of signal assay in a format compatible with microtiter 1536-well quantitative high-throughput screening (qHTS). Continue reading “Genome Editing and Reporter Technologies Enable Endogenous Pathway High-Throughput Assays”

Tips for a Social Media User

Posted on by Sarah Dewbre

Social%20Networks_smallerAs an HR professional, I attend different seminars and conferences to obtain credits for my HR certification. We had a SHRM (Society for Human Resources Management) state conference last week, and I learned all kinds of new strategies involving recruitment, succession planning, employee engagement and change management. One topic was present in every session I was in: social media. How to recruit through social media, engage employees in social media, and how to maintain your company and personal brand through social media.

One session in particular was focused solely on social media, and how it affects the processes of hiring, discipline and discharge. Continue reading “Tips for a Social Media User”

Choosing Your Subcloning Strategy

Posted on by Kelly Grooms

Before you begin your subcloning, you need to know: The restriction enzyme (RE) sites available for subcloning in your parent vector multiple cloning region (or in the insert if you need to digest the insert); the RE sites available in the destination vector multiple cloning region (MCR); and if these same sites also occur in your insert. Once you know this information, you can use the chart below to decide which subcloning strategy to use.

4498MA-[Converted]

To learn more about subcloning, visit our Subcloning Notebook.

Use of Cell-Free Technology to Evaluate Nuclease (TALEN) Activity on Target DNA

Posted on by Gary Kobs
ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;
ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;

Transcriptional activator-like effector nucleases (TALENs) have rapidly become a technique of choice for precision genome engineering. TALENs are custom-designed nucleases that consist of a modular DNA-binding domain fused to a monomeric, C-terminal FokI nuclease domain (1). TALENs work in pairs and are designed to recognize and bind to tandem-oriented sequences in genomic DNA, separated by a short spacer (15–30 bp). TALEN binding causes dimerization and activation of the FokI nuclease domains, which results in cleavage of the DNA within the spacer region. Small insertions or deletions (indels) are frequently introduced at this site, as the result of errors made during DNA repair by nonhomologous end-joining (NHEJ). These indels can be up to several hundred base pairs in length and result in frameshift mutations that lead to the production of truncated or nonfunctional proteins.

Successful use of TALENs for inducing targeted mutations has been reported in many conventional models, for example: mice, Xenopus and D. melanogaster. TALENs are also reported to be functional in a variety of other invertebrate arthropods, including mosquitos,silkworm and cricket. A recent publication (2) illustrates the use of TALEN technology for the genetic manipulation in P. dumerilii (marine ragworm).

Continue reading “Use of Cell-Free Technology to Evaluate Nuclease (TALEN) Activity on Target DNA”

Detecting Inhibition of Protein Interactions in vivo

Posted on by Isobel Maciver
Protein Interactions with NanoBRET

In a paper published in the September 2014 issue of ACS Medicinal Chemistry Letters, researchers from GlaxoSmithKline in the UK and Germany report on the discovery, binding mode and structure:activity relationship of a potent BRPF1 (bromodomain and PHD finger containing protein family) inhibitor. This paper came to our attention as it is one of the first publications to apply Promega NanoBRET technology in an vivo assay that reversibly measures the interaction of protein partners. The technology enabled the identification of a novel inhibitor compound that disrupts the chromatin binding of this relatively unstudied class of bromodomain proteins.

What exactly are bromodomains and why do they matter?
Bromodomains are regions (~100 amino acids) within chromatin regulator proteins that recognize and “read” acetylated lysine residues on histones. These acetylated lysines act as docking stations for regulatory protein complexes via binding of the bromodomain region. Because of their role in chromatin binding and gene regulation, bromodomains have attracted interest as potential targets for anti-cancer treatments. Although some bromodomain-containing proteins (e.g., those in the bromodomain and extraterminal domain (BET) subfamily) are well characterized and have been identified as potential therapeutic targets, others are less well understood.

Continue reading “Detecting Inhibition of Protein Interactions in vivo”

Riboprobes: RNA Probes Are Still Valuable Research Tools

Posted on by Gary Kobs

9613ca[1]Riboprobes are RNA probes that can be produced by in vitro transcription of cloned DNA inserted in a suitable plasmid downstream of a viral promoter.
Viruses code for their own RNA polymerases, which are highly specific for the viral promoters. Using these enzymes, labeled NTPs, and inserts in both forward and reverse orientations, both sense and antisense riboprobes can be generated from a cloned gene.
Transcription of RNA is performed with the appropriate RNA polymerase (T3, T7 or SP6), depending on the RNA polymerase promoter sites present in the chosen vector. Because these polymerases are extremely promoter-specific (i.e., there is almost no transcriptional cross talk), virtually homogeneous RNA can be obtained using plasmid DNA as the template in a transcription reaction. When it is desirable to copy only insert DNA sequences, the plasmid is linearized at an appropriate restriction site before the transcription reaction and only discrete “run-off” transcripts are obtained, virtually free of vector sequences. RNA transcripts may be used to generate radioactive probes for hybridization to Northern and Southern blots, plaque and colony lifts as well as non-radioactive probes (i.e, labeled with digoxgenin)for in situ hybridization.

Recent references using riboprobes include: Continue reading “Riboprobes: RNA Probes Are Still Valuable Research Tools”

Improving Cancer Drug Screening with 3D Cell Culture

Posted on by Sara Klink

Differential contrast image of HCT116 colon cancer spheroid grown in a 96-well hanging-drop platform after seeding with 800 cells. Copyright Promega Corporation.
Differential contrast image of HCT116 colon cancer spheroid grown in a 96-well hanging-drop platform after seeding with 800 cells. Copyright Promega Corporation.
Tissue culture using primary or cultured cell lines has long been a mainstay of testing compounds for inhibiting cell growth or promoting apoptosis during screening for cancer drugs. However, the standard culture conditions result in monolayers of cells, dividing and growing across the bottom of a well, plate or flask in a single layer. The drawback of this technique is that organisms do not come in monolayers; a three-dimensional (3D) spheroid is closer to the in vivo state, especially if the spheroids are made up of more than one cell type like tumors in multicellular organisms. Even more beneficial would be using 3D cultured cells in high-throughput screening to facilitate compound profiling for target effectiveness and cytotoxicity. In a recent PLOS ONE article, researchers used normal and breast cancer cells both in monoculture and coculture to test a set of compounds and found results differed between 2D and 3D cultured cells. Continue reading “Improving Cancer Drug Screening with 3D Cell Culture”