Videos uploaded by user “OriGene Technologies Inc.”
Quick learning of CRISPR/Cas9
What is CRISPR? What is Cas9? How does CRISPR system work? What is needed to perform genome engineering in your lab? This short video introduces the basics of this novel technology, as well as genome editing/knockout products offered by OriGene. For more details, please visit http://www.origene.com/CRISPR-CAS9/ Targeted genome modification has been hampered by the lack of an efficient tool to create a double-stranded break at a designed location. The CRISPR/CAS system is an RNA-guided nuclease system for the targeted introduction of double stranded DNA cleavage. Originally discovered in bacteria as an acquired defense against foreign pathogens, CRISPR/CAS technology has been demonstrated to work successfully in all the common model organisms, making it one of the hottest technology breakthroughs in 2013. Targeted genomic cleavage requires two key elements: a homing device and an endonuclease. In the CRISPR/CAS platform, the homing device is guide RNA, or gRNA and a CAS nuclease. The gRNA contains a twenty-nucleotide target sequence immediately upstream of a Protospacer Adjacent Motif or PAM, linked to a tracrRNA scaffold. This is sufficient to direct the CAS9 nuclease to the complementary site in the genome and create a double-stranded break. Compared to previous genome editing methods, such as ZNF and TALEN, CRIPSR/CAS is cheaper, quicker, and more accessible for researchers. Due to its amazing simplicity, CRIPSR-based genome editing can be achieved with a simple transfection. OriGene offers an all-in-one vector, pCas-Guide, engineered with all the essential elements for targeted cleavage. • The codon-optimized Cas9 expression • A cloning site for a twenty nucleotide target sequence • And a gRNA scaffold downstream of the cloning site to be transcribed into a complete gRNA by a U6 promoter. With a target sequence cloned into this vector, cells transfected with these constructs will express Cas9 nuclease and the guide RNA, which will then lead to a sequence-specific double-stranded break in the genome. Other vector variations are also available to provide added convenience, such as GFP for transfection monitoring or Lenti-backbone for viral delivery. For researchers who perform mRNA microinjection or mRNA transfection, a T7 driven expression vector is available. With the CRISPR/CAS system, genome editing is now smooth sailing. Two types of editing are commonly used. Homologous recombination utilizes a repair template. The desired changes are flanked by left and right homologous arm sequences. Upon double crossover, the desired change is integrated into the genome. Applications include gene knockout, gene-tagging, and site-specific mutagenesis. Non-homologous end joining, where the ends of the break are joined together, introduces random mutations, insertions, and deletions. This can also be used to create variant libraries. Applications for CRISPR/Cas9 are endless. You can generate an in-del variant library at a targeted locus. You can knock out a target gene and simultaneously knock-in a functional cassette, such as a mammalian selection marker or a fluorescent marker. And you can introduce pre-designed mutations. One common application is the promoter study. It requires the insertion of a reporter gene, such as luciferase and/or a GFP, at the first exon of a target gene. This results in knocking out the target gene and knocking in the reporter, therefore enabling the researcher to monitor the promoter activity. For this application, OriGene offers a complete solution. Simply search with a gene symbol, and you can find a fully functional kit containing • Two gRNA vectors, • One donor vector, • And one scramble control vector Another common application is the safe-harbor insertion of transgenes. Compared to other transgene insertion methods, this method provides total control, for insertion site, orientation, as well as for copy number. A ready-to-use kit for this application is coming soon from OriGene as well. A ready-to-use kit for this application is now available from Origene. Origene brings you a complete CRISPR/Cas solution that is simple and easy to use. To learn more please visit the Origene website.
Gene Knockout using CRISPR, a Video Tutorial
CRISPR technology democratized for genome engineering. This game-changing breakthrough makes it feasible for every researcher to perform gene knockout without any major undertaking. To make things even easier, OriGene provides a pre-designed knockout kit for every human and mouse gene. Each kit includes all the plasmid constructs (2 targeting gRNAs, one scramble gRNA and a donor template vector) and a simple protocol. This video tutorial serves as a step-by-step guide for the gene knockout experiment.
Lentivirus production - packaging, titer-measuring and stabilizing
Lentivirus production becomes very easy and simple when lentiviral packaging, lenti titer and lenti concentration kits are available. OriGene is one stop-shop for lenti products from lenti-ORF and lenti-shRNA to lentiviral production kits. All of OriGene's lentiviral particles are provided in the lenti stabilizer solution, which extends lentivirus storage to at least 1 year at -80 Celcius.
OriGene Presents: The Flow Cytometry Protocol
New to flow cytometry? Learn from this 2-min video protocol with step-by-step instructions. OriGene, the largest monoclonal antibody developer, shares the protocol with the scientific community. This is also the same protocol that OriGene uses to validate our TrueMABs and UltraMABs.
Lenti Biosafety
Lentivirus is a powerful gene delivery tool. Lentivirus can delivery genes to both dividing and non-dividing cells. What is the Biosafety level? Is it safe to use lentivirus? What precautions should you follow?
Educational Seminar: Expression Hosts for Recombinant Proteins
This video overviews the pros and cons of various host expression systems for the generation of recombinant proteins. OriGene utilizes human cell lines to produce full-length human proteins: the best strategy to preserve the native structure of a protein, as well as the activity of the protein!
CRISPR Webinar by Drs. Barrangon, Zhang and Church
Webinar sponsored by OriGene and held by GEN, Genetic Engineering & Biotechnology News This webinar on the genome editing system CRISPR/Cas9 is given by three scientists who have been pioneering in the development and application of this technology: Dr. Radolphe Barrangon, North Carolina State University Dr. Feng Zhang, MIT Dr. George Church, Harvard University
PD-L1 IHC Antibodies with  Ultra Specificity & Sensitivity: Tested and Proven
This short video is an introduction on how OriGene has utilized its proprietary UltraMAB and TrueRAB technologies to develop a panel of anti-PD-L1 monoclonal antibodies with superior performance on IHC. For more information about OriGene products and services, please visit http://www.origene.com.
Webinar: Genome Editing with CRISPR/Cas9
OriGene offers CRISPR/Cas9, the latest and effective tool for genome editing. This webinar covers the basics of the CRISPR/Cas9 system, as well as the applications of this novel technology.
Lenti webinar -Why use lenti, biosafety & detailed protocols
Lentiviral vectors are a powerful gene delivery tool; high transfection efficiency, simple and safe to use. Lentivirus can transduce both dividing and non-dividing cells and animals.
Webinar: Gene Knockout with CRISPR
Webinar: Gene Knockout with CRISPR CRISPR/Cas9 technology enables every researcher to do gene knockout at the chromosomal level. CRISPR technology democratized for genome engineering. This game-changing breakthrough makes it feasible for every researcher to perform gene knockout without any major undertaking. To make things even easier, OriGene provides a pre-designed knockout kit for every human and mouse gene. Each kit includes all the plasmid constructs (2 targeting gRNAs, one scramble gRNA and a donor template vector) and a simple protocol. This webinar shows as a step-by-step guide for the gene knockout experiment.
Product Overview: Over-expression Lysates as WB Positive Controls
Antibodies are one of the most popular lab reagents, but their specificity and affinity are rarely confirmed. OriGene provides a collection of over-expression protein lysates to test and validate antibodies. These over-expression protein lysates can be used for antibody validation, as controls in Western blotting, ELISA, and other antibody assays. They can also be applied in the study of protein functions, in immunoprecipitation and reverse phase protein arrays.
How to Make Lentiviral Particles - Recorded Webinar
Makeing lentivirus is a multi-step process, including lentiviral packaging, lentiviral particle enrichment, lentivirus concentration, how to measure lentiviral titier and how to increase lentiviral stability during storage. This webinar provides detailed protocols about how to make high titer virus.
ERCC1 Antibody 4F9, an UltraMAB story on antibody specificity
Antibody specificity is of pivotal importance in clinical diagnosis, OriGene has developed UltraMAB™, a class of monoclonal antibodies of ultra specificity for such rigorous requirement. This video illustrates the importance of using UltraMAB™ in IHC-based clinical diagnosis. Nuclear protein ERCC1 was used as an example. ERCC1 is used as a biomarker to predict caner patient responsiveness to cisplatin chemotherapy, and OriGene discovered that the existing antibody against ERCC1 was not specific. OriGene used its high-density protein microarray to identify and validate a truly specific anti-ERCC1 UltraMAB™, clone 4F9, which independent researchers have shown to be highly specific and which is now recommended for prognostic use.
HuSH 29mer -  Highly Potent shRNA
HuSH-29 shRNA, OriGene's 29mer shRNA, is proven to be more effective than the traditional 21mer shRNA in knocking down gene expression. In addition, 29mer shRNA reduces off-target effects. shRNA plasmids are available in non-tag, GFP and RFP vectors, as well as lenti-viral vectors. All shRNA kits come with 4 gene-specific shRNA constructs and 1 scrambled negative control. To learn more, please visit www.origene.com/shRNA/
Product Overview: OriGene DNA clones
OriGene has the world's largest collection of full-length cDNA clones, as well as cloning vectors and organelle markers. OriGene also provides services of gene synthesis. This video brochure introduces the unique utilities of OriGene clones, and how you will facility your research and development with these products.
Webinar: TrueORF clones - Expression Validated ORF clones
Webinar on OriGene's most prestigious products -- TrueORF clones. Every TrueORF Gold clone is fully sequenced and expression validated. 10ug ready-to-use DNA is delivered with the sequencing chromatogram available online. Visit the OriGene website for more details: http://www.origene.com/ORF/
Lenti webinar -lenti advantages, biosafety & production-June 2018
Lentiviral system is an efficient gene delivery tool. The most recent recorded webinar covers why lentivirus is used to delivery genes, the biosafety and how to produce lentiviral particles.
UltraMAB Antibody: How to Test Antibody Specificity
UltraMABs, the ultra-specific and application-validated antibodies developed for the detection of cancer biomarkers and other important diagnostic targets. This video overviews a breakthrough technology "High-density Protein Microarray" for the examination of antibody specificity, as well as the application of this technology in the generation of UltraMABs.
OriGene Protocol: TissueScan qPCR Arrays
TissueScan™ Cancer and Normal Tissue cDNA Arrays are developed for differential gene expression analysis and validation among hundreds of different human tissues in less than two hours. OriGene also offers comprehensive human cancer and normal tissue products from over 140,000 high quality human tissues. The products include RNA, DNA, Protein and Sections from these tissues. Follow this URL to learn more: http://www.origene.com/qPCR/Tissue-qPCR-Arrays.aspx
Antibody validation using knockout (KO) cell lysates
Many methods have been used by scientists to prove the specificity of an antibody. Out of these methods, Knockout (KO) validation is by far the most trusted validation process for antibody specificity. CRISPR technology has made this validation method more accessible to the antibody developers. This webinar will talk about how OriGene validate the specificity of antibodies through this new exciting technology. For more information, please visit us at www.origene.com.
TrueORF GOLD: the only expression-validated cDNA clones
Expression cDNA clones are an important for gene function study. TrueORF Gold is a class of cDNA clones ideal for protein expression 1) Western blot test of the transfection lysate 2) Sanger sequencing of each base pair 3) in-stock for delivery as transfection-ready plasmids. Why risk your project or grant application? Quality research deserves quality cDNA clone. Trust TrueORF GOLD.
Product Overview: TrueMAB Antibody
OriGene's TrueMAB mouse monoclonal antibodies are generated against native epitopes. Full-length human proteins produced in human cells are used as antigens. Every TrueMAB antibody has been extensively validated for multiple applications, including WB, IHC, IF/ICC, IP and Flow cytometry. Some of them are also tested for ELISA and Luminex as pairs. A positive control is included for FREE with every TrueMAB if applicable. For more details, please visit www.origene.com/Antibody/TrueMAB/
Educational Seminar: UltraMAB, the Ultra-specific Antibody
This video introduces a breakthrough technology -- High-density Protein Microarray -- that can be used to test the specificity of an antibody. It has been proved that some commonly used diagnostic monoclonal antibodies on the market are not specific to their targets. Utilizing this technology and the capacity of monoclonal antibody generation, OriGene has successfully developed the ultra-specific antibody against HER2 for breast cancer, as well as many other cancer biomarkers.
ALK UltraMAB Antibody -  Clone OTI1A4
This video introduces the necessity of having a good ALK antibody for lung cancer diagnostics and showcases is the ALK UltraMAB antibody, clone OTI1A4 is the ideal choice for the IHC application. For detailed validation of this antibody, please visit http://www.origene.com/antibody/ALK-UM800118.aspx
Antibody Specificity and KO Lysates
Antibody specificity has been a well-recognized problem in the research community. OriGene approached the problem using CRISPR to create a long list of Knockout Lysates. Cell lines with both alleles of a specific genes knocked out by CRISPR are called Knockout Cells, or KO Cells. Cell homogenates from the KO cells are the KO cell lysates. OriGene has worked through our own monoclonal antibodies.Briefly the KO cell and its parental lysate were analyzed by western blotting. To qualify for a KO validated antibody, the target protein should be detected in the parental lane, but be absent in the knockout lane. The same blot was stripped and re-probed with a loading control antibody, demonstrating equal loading of the 2 lysates. So far, over 100 of OriGene’s TrueMABs has been validated using this rigorous process. And the list is growing. They are prominently labeled by the KO validated logo. The KO-validated antibody list includes some popular names, such as HER2, IDH1, NOTCH1, c-MET, KI67, and many more. Both the KO cell lysate and the KO validated antibodies are searchable by typing a gene symbol into the search box. Are you concerned with the validity of your antibody experiments? Come to OriGene to select the pre-validated antibodies or choose the KO lysates to validate your own antibodies. For more details, please visit the OriGene website.
Webinar: Over-expression Lysates as WB positive controls
Webinar on over-expression lysates from HEK293T cells -- Why you should include them as positive controls with your Western blot experiements. Visit OriGene website for more details: http://www.origene.com/Lysate/
UltraMAB by OriGene - The Ultra Specific IHC antibodies
UltraMAB ™ is the most specific immunohistochemistry (IHC) antibody in the marketplace. Its specificity has been tested with OriGene's unique high density chip of 10,464 (10K) human proteins. This protein microarray technology has been used to validate the specificity of an existing ERCC1 diagnostic monoclonal antibody, and has been applied as a screening method to identify the most specific UltraMAB monoclonal antibody for diagnostic purposes. Follow the URL to learn more: http://www.origene.com/UltraMAB/
Webinar: New Tool for Antibody Specificity Testing
This webinar introduces a new technology OriGene develops for the validation of antibody specificity. Non-specific antibodies recognize "off-targets" in addition to their intended targets, which could generate false-positive results. This new tool, High-density Protein Microarray, has been used to test the specificity of commercial diagnostic antibodies and found some are not specific to their targets. This tool is also applied to identify the ultra-specific antibodies UltraMABs.
TissueScan: Gene expression Profile  in Cancer Tissues via qPCR
TissueScan™ Cancer and Normal Tissue cDNA Arrays are developed for differential gene expression analysis. It validation among hundreds of different human tissues in less than two hours. Tissue cDNAs of each array are normalized first-strand cDNA from clinical tissues. Each cDNA sample is synthesized from high quality total RNAs of pathologist-verified tissues, normalized and validated with beta-actin in two sequential qPCR analyses, and provided with clinical information and QC data. Learn from this video on how to survey gene expression level of any potential biomarkers among a full-spectrum of cancer tissues in 2-hrs. For more information, please visit http://www.origene.com/qPCR/Tissue-qPCR-Arrays.aspx
OriGene Webinar Series: TissueScan cDNA Arrays
Webinar on TissueScan products -- Cancer and Normal Tissue cDNA Arrays -- how you obtain qPCR gene expression results of hundreds of human samples in 2 hours. Follow this link for more details: http://www.origene.com/qPCR/Tissue-qPCR-Arrays.aspx
TissueFocus: Normal and Cancer Human Tissues & Derivatives
Over 140,000 human tissue samples collected from U.S. medical centers using the industry's most strict ethical requirements and IRB protocols. OriGene offers normal and cancer tissue slides from these samples. In addition, tissue derivative products are also provided, including total RNA, genomic DNA, and tissue protein lysate. This video introduces how to use the search tool on OriGene website to find the appropriate tissue products for your research. Visit http://www.origene.com/tissue/tissueSearch.aspx for details.