Site-specific large fragment DNA knock-in rat model: TARGATT(TM)- Rat Models

For decades, mice have been the major transgenic in vivo model for gene function studies and drug discovery. Their low maintenance cost and easy manipulation made them a preferred tool for pre-clinical research. However, as the need for a better understanding of human genetics has grown, there has been an increased demand for improved transgenic in vivo models. Site-specific, transgenic rats are gaining importance as a preferred in vivo model for researchers who seek a better representation of human genetics and physiology.  Their similarity to human physiology and pathology makes them a better in vivo model than mice for the study of cardiovascular, diabetes, neurological, psychiatric, and autoimmune dysfunctions.

Major improvements in site-directed mutagenesis using techniques such as TARGATTTM, CRISPR/Cas9, ZFNs and TALENs have made the generation of transgenic rat models possible. Applied StemCell’s proprietary TARGATTTM technology enables the insertion of any gene of interest (up to 20 kb) with high efficiency and guaranteed expression. The gene is integrated into a well-characterized, transcriptionally active locus in the rat genome engineered with an attP docking site The DNA is injected directly into the rat zygote, thereby bypassing rat embryonic stem cells (rES). Our innovative technology can generate site-specific, single copy knock-in rat in 4 months.

We recently presented the TARGATTTM-rat model and related services at the prestigious 13th Transgenic Technology Meeting held in Prague, Czech Republic from March 20-13, 2016. Download a copy of the poster

To learn more about the site-specific knock-in TARGATTTM-Rat Model, Register for our upcoming webinar.

Integrase-based TARGATTTM Method for Generating Site-Specific Transgenic Rat Models

Date: Tuesday, April 12, 2016

Time: 11:00 am – 12:00 pm

Presented by: Dr. Ruby Yanru Chen-Tsai (CSO, Senior VP, R&D), Applied StemCell, Inc.

The novel TARGATTTM technology and the complementary CRISPR/Cas9 system provide a versatile gene editing tool set for the generation of:

For more details, and to explore our other services and products, please visit us at www.appliedstemcell.com or contact us at info@appliedstemcell.com.

CRISPR-Cas9 Screening Reveals Novel Therapeutic Targets for Cancer Therapy

CRISPR-Cas9 Screening: Genome-wide screens that employ CRISPR-Cas9 are playing a key role in the identification of proteins and pathways that play an integral role in disease. In this study, the authors use CRISPR-Cas9 screening to identify drug targets for cancer therapeutics by screening an array of protein functional domains. Specifically, by screening of 192 chromatin regulatory domains in murine acute myeloid leukemia cells, the authors were able to identify six known drug targets and an additional 19 potential target candidates.

Notably, the authors found that targeting gRNAs to exons that encode essential protein domains is a preferable strategy to targeting the 5’ exons of candidate genes. This strategy enabled stronger phenotypes, and the detection of gene dependencies that may otherwise go unidentified using 5’ exon targeting approaches.

Shi et al. Discovery of cancer drug targets by CRISPR-Cas9 screening of protein domains. Nature Biotechnology. 2015, 33: 661-667. DOI: 10.1038/nbt.3235

 

in vivo Genome Editing with CRISPR-Cas9 Enables Elucidation of a Therapeutic Mediator in Epigenetic Treatment for Pancreatic Cancer

in vivo genome Editing with CRISPR-Cas9.

Pancreatic ductal adenocarcinoma is a lethal cancer with a median survival of 6 months, and a 5-year survival of less than 5%. Despite significant advances in the detection and treatment of other cancers, the prognosis for pancreatic cancer has shown little improvement over the past 40 years.

Reporting in Nature Medicine, Pawel Mazur and colleagues evaluate combined inhibition of BET (bromodomain and extraterminal) family proteins and histone deacetylases (HDACs) as epigenetic targets for the treatment of pancreatic cancer. The authors find that combined treatment with SAHA, a HDAC inhibitor, and JQ1, an inhibitor of BET proteins, is able to substantially reduce tumor burden in a mouse model of the disease, resulting in a significant improvement in overall survival.

After identifying p57 (or Cdkn1c) as a potential mediator of the therapeutic effect, the authors utilized direct injection of lentivirus into the mouse pancreas for in vivo p57 knockout using CRISPR-Cas9. Tumors with successful p57 deletion showed a reduction in apoptosis upon treatment with SAHA and JQ1, thereby supporting potential role of p57 in mediating the therapeutic response.

The study elucidates a potential epigenetic susceptibility of pancreatic cancer, providing an opportunity for new therapeutic interventions.

Mazur et al. Combined inhibition of BET family proteins and histone deacetylases as a potential epigenetics-based therapy for pancreatic ductal adenocarcinoma. Nature Medicine. 2015, 21: 1163-1171. DOI: 10.1038/nm.3952

CRISPR-Cas9 Cancer – The CRISPR System is Transforming Biology

CRISPR-Cas9 and Cancer. The discovery of CRISPR-Cas9, and the demonstration of its utility for human genome editing, is rapidly advancing the field of genetic engineering. Such a platform offers enormous potential for the study of cancer – a disease that is characterized by, and arises as a result of, many genetic and epigenetic changes. In this Nature Reviews Cancer article, Francisco J. Sánchez-Rivera and Tyler Jacks provide an overview of the ways in which the CRISPR-Cas9 system is enabling discoveries in cancer biology.

Cas9 applications include:

  1. Indel formation and precise genome editing by NHEJ and HDR, respectively
  2. Utilization of CRISPR technology for the generation of germline and non-germline genetically engineered animal models
  3. In vivo and ex vivo engineering of somatic cells for cancer modeling
  4. dCas9-effector fusions for CRISPR-mediated gene activation or repression
  5. High-throughput genetic screens using CRISPR-Cas9 gRNA libraries

The CRISPR-Cas9 system has greatly expanded the available toolbox for scientific discovery in cancer biology. Learn more about the CRISPR-Cas9 cancer services and products that are offered at Applied StemCell.

Francisco J. Sánchez-Rivera and Tyler Jack. Applications of the CRISPR-Cas9 system in cancer biology. Nature Reviews Cancer. 2015, 15: 387-395. DOI: 10.1038/nrc3950

MyEZGel™ Advanced Peptide Hydrogels and 2D/3D Cell Cultures for Cancer Cell Lines and in vivo applications

No More Icing – MyEZGel™ Advanced Peptide Hydrogels and 2D/3D Cell Cultures for Cancer Cell Lines and in vivo applications

Wed, 11/11/2015 11AM – Noon (PST)

  • Existing 3D cell culture tools
  • Methods for MyEZGel™ cell encapsulation, isolation, fixation, and imaging procedures.
  • Case studies using MyEZGel™ for 2D/3D culture of cancer cells, and injection of cells and viruses in vivo. 

(45 min., 35 min. presentation + 10 min. FAQ)

Speaker: Xiuzhi Susan Sun, PhD., Kansas State University

Registration:  https://attendee.gotowebinar.com/register/8961398367836353537

Life science research has been limited by the traditional 2D cell culture system. Scientists are switching to 3D cell culture systems with the hope of more accurately mimicking the native extracellular microenvironment. This webinar will discuss existing 3D cell culture tools, particularly the new peptide hydrogel (MyEZGel™) that is biocompatible and easy to use, and has similar fiber dimension and pore size to the native extracellular matrix (ECM). MyEZGel™ has many unique properties for 3D cell cultures and potential medical applications, such as cell therapy, tissue engineering, and sustained release agents, to name a few. This webinar will present methods for cell encapsulation, isolation, fixation, and imaging procedures. We will also provide a few case studies using MyEZGel™ for 2D/3D culture of cancer cells, and injection of cells and viruses in vivo.

Speaker: Dr. Xiuzhi Susan Sun
Dr. Xiuzhi Susan Sun is a University Distinguished Professor at Kansas State University (KSU) and member of the National Academy of Inventors. Dr. Sun is the primary inventor of a new peptide hydrogel technology with unique features for life science and medical research. Dr. Sun is the founder of the Biomaterials and Technology Lab at KSU. Dr. Sun has strong biomaterials research experience, specializing in protein and lipid structure and functional properties as monomers, polymers, and peptide hydrogel structures for medical applications, biobased adhesives, resins, and coatings for environmentally safe materials. Susan has supervised $20+ million grants to support her research programs in the past two decades. Dr. Sun has published 165 reference articles and holds 11 patents with 6 patents pending. As a productive inventor, Susan closely works with many industrial collaborators. She has successfully commercialized some of her patented technologies, and is a founder of the PepGel LLC Company. Dr. Sun received a Lifetime Achievement Award from the Bioenvironmental Polymer Society.

New Products! 2D3D Cell Culture MyEZGel™ for Cancer Cell Lines

RECEIVE A FREE SAMPLE! Email info@appliedstemcell.com

MyEZGel™ is a protein based solubilized basement membrane synthesized from commonly used amino acids, and minerals at the concentration similar to less than those in the cell media. MyEZGel is an effective scaffold for three dimensional (3D) cell cultures of various cells, particularly for various cancer cells, liver cells, fibroblast cells, skin cells, endothelial cells, and other cell types.  MyEZGel can be used for several types of tumor cell invasion assays, sustained release agent, vaccine adjuvant uses, and for delivery of pluripotent or adult stem cells by injecting into mice for in vivo differentiation functions.

Technology Comparison

2D cell culture: MCF-7 (breast cancer cells) invasion in gels fom the top surface

3D cell culture image with MCF-7

MCF-7 cells in MyEZGel grow for 5 days in 3D in 3mM (0.51% w/v)  hydrogel media. Image shows MCF-7 cells form spheroid shape colonies. Figure from: Hongzhou Huang et al., Peptide hydrogelation and cell encapsulation for 3D culture of MCF-7 breast cancer cells, PLos ONE 8(3): e59482.doi:10.1371/journal.pone.0059482 (2013).

Sample Request Form: info@appliedstemcell.com

 

Applied StemCell Genome Editing Webinar

Applied StemCell is excited to announce our new webinar called: “Choosing the Right Genome Editing Technology: CRISPR, TARGATT™, and beyond.” This webinar is focused on genome editing, and is a fantastic opportunity to learn more about how Applied StemCell can aid you in your research.

The webinar will be hosted on GoToMeeting, and take place on August 25th Tuesday, from 11AM. The entire webinar will take around 50 minutes to complete.

To sign up for our genome editing seminar, please visit: https://attendee.gotowebinar.com/register/7076934392323758593

 

 

 

 

NIH Research Festival Exhibit

NIH Research Festival Exhibit
September 17-18, 2015 • National Institutes of Health • Bethesda, MD

Sponsored by the Technical Sales Association

Location:  Exhibit Tent on Parking Lot 10H at the Valet Entrance to Building 10 on the south side.  

Booth # 858

Exhibit Hours: 

  • Thursday, September 17: 9:30 AM – 3:30 PM
  • Friday, September 18:   9:30 AM – 2:30 PM

http://researchfestival.nih.gov/

CRISPR Rat (SD Strain) Model Generation Service Promotion

Thank you for GROWING with us!

SPECIAL PRICE PROMO: CRISPR Rat (SD Strain) Model Generation Service 

We’re expanding our facility space by more than double to accommodate our growth. We are currently running a BIG SPECIAL THANK-YOU PROMO for CRISPR Rat (SD Strain) model generation services!! Please feel free to contact us for details.

1-866-497-4180

info@appliedstemcell.com

Thank You to Our Transgenic Community.

Applied StemCell would like to extend a thank you to our transgenic community. We are expanding our business & facility!

We’re expanding our facility space by more than double to accommodate our growth. We will close our office Friday, August 28th, 2015 to begin the moving process and will re-open at new location Monday, August 31st at 8:30 a.m. Our business, including website, email, and telephones, will be fully operational during the moving process and you will not experience any changes or delays in service, production, or shipping during that time.

Please update your records with our new contact information:

Applied StemCell, Inc. 521 Cottonwood Drive, Suite 111, Milpitas, CA 95035

Tel: 408-773-8007 Fax: 408-773-8238