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The TransMission

Transfection Blog Delivered by Mirus

Recent Posts

 

Targeting STMN2 Splicing to Treat ALS

May 30, 2023

Today’s TransMission is a SNiP (Small News in Pieces) about stathmin-2, which is encoded by the STMN2 gene. No, this is not a pun on a Jason Statham action movie. STMN2 is a potential drug target for amyotrophic lateral sclerosis (ALS). In this SNiP, we highlight how Baughn & Melamed et al. modulate splicing of STMN2 mRNA with antisense oligonucleotide- and CRISPR-based approaches. The strategies and tools introduced in their study will be useful in the development of gene therapies to treat ALS and related neurodegenerative diseases.

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Media Adaptation for Suspension Cells

May 16, 2023

The success of an experiment can rely on the type of cell culture media used. In certain cases, you may need to adapt your current cell culture to a new medium. A stepwise process may be necessary to minimize stress and to retain normal cellular phenotype. Check out this Tip from the Bench to learn how to gradually adapt suspension cells to a new media formulation.

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How can I increase my viral titers and improve the percentage of full AAVs?

May 2, 2023

In today’s TransMission, we answer the FAQ: “How can I increase my viral titers and improve the percentage of full AAVs?”

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Hitting HIV with Combo Therapy

April 25, 2023

The field of HIV/AIDS medicine is no stranger to combination therapy. In this SNiP (Small News in Pieces), we highlight a paper from Usero et al. that suggests using an mRNA-based HIV vaccine in combination with immunomodulators to clear HIV infection.

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Transfecting Insect Cells, Pt. 3

April 11, 2023

Howdy everyone! Dr. Bees here. So far, we have discussed common insect cell lines and the insect-specific transfection reagent TransIT®-Insect (Part 1) and producing baculovirus with flashBAC™ (Part 2). Today in Part 3, we will do a quick flyover of the pOET Transfer Plasmids, which complement flashBAC™ DNA like bees and honey!

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How is transfection critical to the success of a cell or gene therapy?

April 3, 2023

In this short post, we highlight the importance of transfection for cell and gene therapy.

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What media can I use to form transfection complexes?

March 20, 2023

In today’s TransMission, we answer the FAQ: “What media can I use to form transfection complexes?”

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Zinc Finger Transfection

March 9, 2023

Today's TransMission is a SNiP (Small News in Pieces) about zinc fingers and the subtleties in zinc finger design. Zinc fingers are an alluring biotherapeutic modality because of their natural prevalence in humans, modularity, small size (~30 amino acids per finger) and DNA-targeting ability. However, zinc finger protein design remains challenging because DNA binding specificity and affinity is affected by the presence of adjacent zinc fingers in an unclear pattern. Read today’s TransMission to learn how Ichikawa et al. applied machine learning in hopes of developing a proverbial 'Zinc Finger Rosetta Stone' for designing zinc fingers capable of targeting any genomic sequence.

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Transfecting Insect Cells, Pt. 2

March 1, 2023

Howdy everyone! Dr. Bees is back with the 2nd installment of our insect transfection series, and bee-lieve me you will enjoy this one. Last time, we covered commonly transfected insect cell lines. Today we are going to focus on flashBAC™--what it is and how to use it for producing baculovirus.

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CRISPR’ing Wheat with a Gene Gun

February 22, 2023

What happens when chemical transfection meets gene guns? In today’s Small News in Pieces (SNiP), we highlight biolistic transfection and a study that incorporated TransIT®-2020 Transfection Reagent to genetically modify wheat plants.

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Transfecting Insect Cells, Pt. 1

February 14, 2023

Howdy everyone! Guest writing for the TransMission is one of our field application specialists, Dr. Bees… which is a nom-de-plume, of course! Starting the month of February for you love bugs and valentines, Dr. Bees will be doing a series of insect-themed posts, including transfection solutions that Mirus has developed for insect cells. In Part 1, we begin by introducing the cast of characters and the critical cell biology of insects.

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Regulate pLIVE® Expression Level

February 6, 2023

pLIVE® is a vector system designed for long-term expression in the livers of laboratory rodents. In this Tips from the Bench, we share how gene expression levels can be modulated by changing the relative position of the gene of interest within the pLIVE® multiple cloning site. We also discuss testing the pLIVE® construct in vitro before beginning in vivo studies.

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Transfection Efficiency

January 26, 2023

“How successful was your transfection experiment?” Transfection efficiency is a measure used to answer this question. Because success can be defined in multiple ways, how to calculate transfection efficiency will depend on the type of transfection experiment. In this TransMission Transfection 101 we will cover: the many meanings of transfection efficiency, common reporter assays to measure transfection efficiency, and how to improve transfection efficiency.

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Adapt Adherent CHO Cells to Suspension Culture

January 19, 2023

Chinese Hamster Ovary (CHO) cells are a popular workhorse for production of recombinant therapeutic proteins. Amenable to both adherent and suspension culture, CHO cells offer versatility in growth platform that is favorable to the end user’s requirements. In this Tips from the Bench, we highlight factors that differentiate adherent and suspension CHO cell cultures. Also, we describe a simple protocol for transitioning adherent CHO cells to suspension growth (and serum-free medium, if required).

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Current FDA Approved Gene and Cell Therapies

January 10, 2023

Let’s ring in the new year with a list of all current FDA approved gene and cell therapies to date! Read on to learn more about the 27 different treatments and cures.

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Understand Viral Titer & Quality

December 21, 2022

The cell and gene therapy field has burgeoned in the last decade with drug candidate treatments for many different diseases. Determining the correct patient dosage has become more important than ever to extend the reach of the multi-million-dollar manufacturing runs required to generate these viral therapies. Genome copies (GC), capsids, GC/capsid ratios, percent full, functional titering… what does it all mean? In this Transfection 101, we summarize commonly used approaches to verify that manufactured viral vectors are of high quality and functionality, appropriate for use in patients.

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Storage of TransITs

December 6, 2022

Proper storage and handling of TransIT® reagents are required to maintain optimal transfection performance. This is a quick reference page for storage and handling of Mirus TransIT® Transfection Reagents and Kits.

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Thank You

November 23, 2022

Special TransMission incoming!

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Peering into the Proteome of SMA

November 15, 2022

SNiP-ing a recent study related to Spinal Muscular Atrophy (SMA). Digest this SNiP to learn about SMA and the proteomic analysis conducted by Brown et al. to identify biomarkers and determinants of SMA severity.

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Ancillary Materials

November 3, 2022

For those close to the cell and gene therapy world, the term ‘ancillary material' may be as familiar as ‘DNA.’ However, if you are navigating the regulatory landscape of cell and gene therapy manufacturing for the first time, understanding this terminology and its importance may be more of a challenge. Learn more in this TransMission Transfection 101!

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Toxic Transfections

October 25, 2022

Why do cells die after transfection? How can you measure cytotoxicity? These questions answered in this TransMission Transfection 101, where we tackle cytotoxicity. Read on for advice on how to reduce transfection-related toxicity.

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Leveraging Avian AAV in Pekin Duck Research

October 14, 2022

Though typically seen in reference to human disease, AAV has applications apart from therapeutics for otherwise untreatable maladies. Today we SNiP a proof-of-concept study that features a unique use for TransIT-VirusGEN® in production of avian AAV for the transduction of Pekin duck cells in vitro and in vivo.

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Transfection Troubleshooting Checklist

October 4, 2022

Having trouble with your transfection experiment? Go through this transfection troubleshooting checklist to eliminate potential transfection problems.

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Which reagent should I use for transfecting oligos?

September 21, 2022

In today’s TransMission, we answer the FAQ: “Which reagent should I use for transfecting oligos?”

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SNiP-ing Success with CRISPR

September 6, 2022

It’s fitting that we SNiP (Small News in Pieces) an article about CRISPR. Read on if you think your CRISPR experiments are plagued by re-cutting, which can decrease gene-editing efficiency and introduce undesired mutations. Co-transfection with hideRNA may be a quick fix!

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Great Cycle Challenge

September 2, 2022

Special TransMission incoming from our GCC team leader (a.k.a. Director of Chemistry at Mirus Bio) Nick Rossi, Ph.D.!

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Optimizing siRNA Delivery

August 24, 2022

Because siRNA differs in size and structure from plasmid DNA, transfection reagents can be optimized and formulated differently for delivery of these RNA molecules. After selection of the appropriate transfection reagent, optimization of several experimental parameters is key to achieving the highest efficiency siRNA transfection and corresponding effective knockdown of target gene expression. Here we highlight our top "Tips from the Bench" for optimizing siRNA delivery. Note: most of these tips are applicable to delivery of other classes of oligos, including but not limited to miRNA and antisense oligonucleotides.

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Zap That!

August 8, 2022

We SNiP a study by Willerslev-Olsen et al., featuring Ingenio® electroporation of T cells with siRNA.

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Tell Me More About Telomeres

July 26, 2022

Today's TransMission is a SNiP (Small News in Pieces). Read how Jack et al. use TransIT® and Label IT® to study telomeres.

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Optimizing Plasmid DNA Delivery

July 13, 2022

Setting up successful transfection experiments for the first time can be a daunting task, especially when working with stubborn cells that are refractory to transfection. Variables such as cell line, passage number, confluency, DNA quality and even transfection reagent formulation all contribute to the likelihood of success (or failure) of a given transfection experiment. Here, we highlight our top "Tips from the Bench" for optimizing plasmid DNA delivery.

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Extending Viability in CHO Cells

June 30, 2022

A common question we encounter is: how can I extend CHO cell viability through the process of protein production? One common practice to maintain higher viability is to shift cell cultures to mild hypothermic conditions, e.g. 37°C to 32°C. Here, we discuss a hot (er… cold?) tip involving the CHOgro® High Yield Expression System, which also supports additional flexible protocol options for researchers who cannot shift cultures to 32°C or who desire extended cellular viability beyond one week following transfection.

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Deliverance: understanding transfection complexes

June 15, 2022

How do Mirus transfection reagents deliver? Today we'll discuss what transfection complexes are and how nucleic acids are transfected into cells. This post will explain the purpose of steps in transfection protocols, including: transfection complex incubation, amount or ratio of reagent to nucleic acid to add, and what media to use for transfection complexes.

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From AAV to TAZ

June 6, 2022

Today's TransMission is a SNiP (Small News in Pieces). Read how Hwang et al. use AAV to study mitochondrial biogenesis.

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(Re)Starting the Conversation

June 1, 2022

In this inaugural post, we introduce the goals of and Technical Support team behind the Mirus Bio Blog. A brief timeline of some milestones in Mirus history is also presented.

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