New product of the week 01-11-10 to 01-17-10

Light-driven proton pumps were used to to silence the activity of genetically specified neurons in a report by Chow et al. in the latest issue of Nature. Genetic delivery of proton pump proteins, Arch and Mac, will be of significant help to neurology labs. We have started put these genes into Allele Biotech’s ready-to-infect lentivirus and retrovirus, which will be available for shipping within a week or two. Allele Biotech decided to list these viral particle products as this WEEK’S NEW PRODUCT among a good number of choices to reflect our model of doing business and science: move fast and stay on the front edge of multiple moving fields.

New product of the week 01-11 to 01-17-10: arch and mac Expression Lentiviral Particles Cat # ABP-RP-TLCARC or TLCMAC.

Promotion of the week: 20% off our most popular TA cloning kit with top of the line competent cells, as announced every Monday through our social networks. Follow us there is you want to get them in time to make a purchase at the deeply discounted prices.

Tags: arch, channelrhodopsins, Chow et al., Halorubrum sodomense, ion modulator, Light-driven, mac, optogenetic, proton pumps, silence of neurons, silencing of neural activity

GPCR: problems and resolutions in high content screening, Part II

2) Without a mark (continued from Part I by Niels Yuhui Ni, MD Ph.D. of Allele Biotech)

In this section, the author focused on the “Label-Free” system. “A newer alternative that has given hope” because these systems are closer to the real cellular conditions most GPCR studies are meant to address. However, label-free systems must depend on complicated detection system for high content analysis. As commented, “For some scientist though, this technology is simply still too new and for now, too expensive for many categorical assessments…” While I am a believer in label-free detection, however, here is my question, for this group of scientists, are there any good alternatives now, before the label-free systems become more accessible? Using the traditional over-expression cell lines seems less and less attractive (see Part I under the same blog topic here)? A system that combines immortalized primary cells and a non-integrated expression system could be a nice system that does not require high end equipment or heavy commitment in technology development especially if the components were commercially available. The third, critical component of this system could be the application of newer, brighter and monomeric and thus less toxic fluorescent proteins or FRET pairs as sensors. Obviously part of the reason that I thought about such a system was because our research team at Allele Biotech has the background in all 3 components, whereas others might have their own preferred methods. To me, it seems that immortalized primary cells present a renewable cell source, and for non-integration delivery, we prefer Baculo viral delivery vehicles for mammalian infection (called BacMam by some). Both platforms are being offered as products or services already or in the pipeline teed up for launching.

Baculo2Mam non-intergrated viral delivery system:
Many people may know about Baculovirus such as in the Bac-to-Bac system from Invotrogen or the Sapphire baculovirus system from Orbigen (acquired by Allele Biotech). But how many of us know that even though mammalian cells are not the nature host of baculovirus, they still can be infected with modest modifications on the virus. Both the safety and efficiency of Baculovirus for mammalian use are superb. Based on the data from our customers’ projects, most protein expression require baculoviral protein expression in insect cells, only about 10% require Baculo2Mam. We actually feel a sense of responsibility for introducing this technology to as many as researchers as possible.

The following list shows some of the advantages of Baculo2Mam I can list right now, for more details check back on our blog articles in coming days or contact us at any time for discussion.
1) Baculoviruses are Risk Group 1 or biosafety 1 agents.
They are produced in insect cells and can not replicate in mammalian cells. They express genes in human or mouse cells in non-integrated state for about 2 weeks (varies in different cells).
2) Baculoviruses can be easily generated in high titer and production rapidly scaled-up.
That is when compared with other viral systems. For example, baculovirus is a budding virus that is released into cell medium, unlike adenovirus that requires lysing cells during productions. Allele Biotech now provides Baculo2Mam viral packaging service at an affordable price for routine use. Your viral clones can be stored in Allele Biotech’s Baculo2Mam virus bank; if you need the virus again, you can just order a production service at an even lower price.
3) Broad host cell range including many primary cells.
Many terminally differentiated primary cells such as neuron, adipocytes have been tested in Allele Biotech’s lab as target for modified Boculovirus. To assess the infection efficiency, you can order a pre-made Baculo2Mam-mWasabi GFP or Baculo2Mam-LanRFP control for a test run. Once you order custom or regular Baculo2Mam products, the cost of the control will be credited back.
4) Up to now, little or no cytopathic effects were observed of using baculovirus in mammalian cell cultures.
5) Other points that may be related to GPCR assays in relevance to mimicking natural cellular environment:
a) Delivery of biosensor to cells just prior to assay without establishing cell lines
b) Large insert capacity for expressing long cDNAs.
c) Multiple virus transductions, simultaneous delivery of multiple genes
d) Expression level can be adjusted by viral titer
e) Finally, Baculo2Mam Viruses can be stably stored at 4oC for up to 3 months, and even longer as seed stocks (i.e. titer will drop but still amplifiable).

Promotion of the first week of 2010:

in the spirit of celebrating Allele’s 10th anniversary and in line with the ongoing “get oligos free for a month” program, we offer $20 off for oligos on 3’ TAMRA or FAM modifications.

New product of the week:

iPS specific gene promoter-fluorescent protein reporter lentiviruses.

Tags: bacmam, baculovirus for mammalian cells, biosensors, cell based assay, cell imortalization service, Fluorescent proteins, GPCR, GPCR assays, immortalizaed primary cells, label-free detection, membrane based assay, viral packaging service

10 Years of Allele Biotech

Facts about Allele’s 10 years in business:

Products

New product lines added in 2009: iPS cells, Camelid Antibodies, DNA synthesis chemicals, Recombinant Proteins

Highlights: HiTiter Lentiviral Systems, Baculovirus for Mammalian Expression (BacMam), Feeder Cells, shRNA on Viral Vectors, shRNA Validation FP Vector, ProperFold Protein Folding Vector, Validated AllHPLC synthetic siRNA

New Service Groups in 2009: Viral Packaging, RNAi Validation/Screening, FP-based Assay Development

Numbers

Since April, we have added at least one new product every week! We currently run one new promotion per week as well.

A bit of history–did you know that…

Allele Biotech obtained 5 NIH grants in its first three years since establishment. As a matter of fact, Allele Biotech was funded entirely by NIH grants

Allele filed its first patent application in its second year of operation, which was on DNA-driven RNAi and resulted in an outlicensing deal with Promega. As result of the applications, Allele has received 3 US patents on DNA-encoded shRNA, siRNA using promoters such as U6 and H1.

During the past 10 years, Allele was the first to sell U6-based RNAi vectors, the only supplier of bFGF-expressing feeder cells for iPSC, most likely a top 3 provider of baculovirus expression systems, camelid antibody products, iPS creating viral particles, and the most active commercial developer of fluorescent proteins.

Tags: 10 years in business, Allele Anniversary, camelid antibodies, NIH grants, oligos, RNAi

GPCR: problems and resolutions in hihg content screening

A review in the December, 2009 issue of Nature Methods, “GPCR: insane in the membrane” by Michael Eisenstein impressed me greatly. It presents a clear and wide view of the GPCR research fields, from the basic background knowledge, to difficulties and current solutions in real cases, to future development. After careful reading and thinking, I felt that there were still several questions left to for me to pounder.

1) The Plot Thickens

“In many cases, you identify a GPCR as a target based on physiological data, and the receptor might be expressed in the brain in a particular neuron, but then you perform a high-throughput screen in over expression, immortalized cell lines that are nothing like the cell in which the receptor normally resides,”This is the inherent problem for all the engineered and artificial cell based assays most widely used in GPCR research right now. As stated in this review, “There are so many ways to be misled by using an over-expressed receptor in a non-native cell line”, it was shocking to me, making me realize that cell line screening does not only have a problem of getting raw data of low quality, but also a problem of being seriously misleading, a problem of direction rather than efficiency.

Directed differentiation of iPS cells and ESCs might be one of the potential solutions to this problem. However, from our experience in iPS cells dedifferentiation and differentiation at Allele Biotech, as a main vendor of iPS cell reagents, it is hard to make this platform in current situation suitable for high content analysis of GPCR. As much as we wish and believe, like many other researchers and the general society, that stem cells will contribute significantly also to the drug screening field, much work needs to be done before that happens.

Primary cells could be a good alternative. Based on data from Allele Biotech’s trade partners in the primary cell business, the primary cells market has been expanding dramatically since late last year. I believe that the utility and advantage of primary cells have been increasingly appreciated. However, there are still some problems with using primary cells in GPCR studies. For example, neurons are hard to obtain and can not amplify in vitro, in which cases there seems to be no chance to satisfy the need by screening. In fact, neuron research is one of the main GPCR research fields. Therefore, this is a must-fix problem.

How about “Immortalized Primary Cells”? The immortalized primary cells maintain the properties of primary cells well, relative to established cell lines. We have built our own advanced immortalization technology, based on which new products and services will be released in the first season of next year. During the last several years, we have successfully immortalized immunological cells and cancer cells. WE have yet to have the experience of working with neurons. Anyone who has interests of cell immortalization is welcome to contact us for collaboration or custom service. “Introducing Cost Effectiveness to your research”, just as the slogan of Allele Biotech, we will be proud to serve you with cutting-edge technology and cost effectiveness!

2) Without a mark to be continued…

By Niels Yuhui Ni, researcher at Allelle Biotech

Allele Biotech is 10 Years Old and Celebrating with FREE Oligos!

Allele Biotech is 10 years old!!!!!!! December 1999 was when Dr. Jiwu Wang and colleagues started this great company as a DNA oligo and siRNA service provider to the San Diego area. Since then he has fearlessly lead Allele into the forefront of the biotechnology industry with multiple RNAi patents, numerous NIH grants, revolutionary iPSC and fluorescent protein technologies, the acquisition of Orbigen, a continuously growing catalog of over 1000 molecular biology products and signature Allele Biotech Reagents, and a business culture that is approachable, encouraging, and reverential of research advancement through global communication and collaboration.

What a difference a decade makes. Today Allele Biotech is a top oligo service provider all around the country with customers in all of America’s major academic institutions. Currently, we are the sole oligo provider on the University of California San Diego’s central purchasing site, Marketplace; a collaboration designed to provide top quality oligonucleotides to UCSD research departments while saving them thousands of dollars annually. We now produce many of our own oligo synthesis and modification reagents, further cutting the costs to our valuable customers which enabled us to continue operations this last year without raising prices due to the worldwide acetonitrile shortage which more than quadrupled in cost! Allele Biotech has stood out over the years not only to our loyal customers but other oligo providers as well. Less than 5 years into our operation we were approached by one of the well known, top three, oligo providers in an attempt to buy us out! We resisted and are still here to proudly serve the research community with the Allele brand!

It all started with oligos…Now ten years later we want to honor our accomplishment by giving away a FREE month of oligos to one lucky customer! To enter you must be an Allele Biotech Facebook fan or friend. A winner will be randomly selected from our friends/fan pool on Sunday, February 14th, 1210 at noon. That lucky winner will receive FREE oligos for the month of March 2010! Limitations apply. Click here for terms and conditions.

Tags: 10 year with Allele Biotech, 10th anniversary promotion, anniversary promotion, FREE oligos, oligo services