• Custom Vector Construction and Modification

    Creative Biogene is one of the leading biotechnology companies which has extensive expertise and experience in providing custom vector construction and modification service for customers worldwide. Creative Biogene's sophisticated equipments, advanced technologies and highly experienced staffs are available to provide you with the complete custom vector construction and modification service, including sequences analysis, gene synthesis & cloning, DNA fragment assembling, DNA sequencing, site directed mutagenesis, and plasmid production, etc. Using fast and efficient construction and modification methods, Creative Biogene can quickly create novel custom vector constructs for large or small projects.
     

    In molecular cloning, a vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell, where it can be replicated and/or expressed (e.g.- plasmid, cosmid, Lambda phages). The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Among these, the most commonly used vectors are plasmids. Nowadays, a wide variety of plasmid vectors are commercially available. However, in some cases, the commercial vectors cannot best match researcher's specific needs and researchers have to rely on in house vector modification.

    With years of development, Creative Biogene is able to provide custom vector construction and modification service to generate custom specialized plasmid vectors with distinct characteristics quickly and reliably. Creative Biogene has developed an advanced gene synthesis platform to modular construct custom plasmid vectors. We obtain the vector modules either by PCR amplification or synthetical methods and using complex cloning strategies when needed to develop completely customized vectors to best fit your unique requirements. Creative Biogene's goal is to provide you with the most affordable, high-quality custom vector construction and modification service to ensure your satisfaction in a timely and professional manner.

  • circRNA-miRNA/RBP Interaction Analysis

    Creative Biogene has developed a complete workflow for your circRNA-miRNA or circRNA-RNA banding protein (RBP) interaction analysis, which has been enhanced with the latest technologies. We offer affordable and high-quality circRNA interaction analysis services for customers. From target miRNA discovery to confirmation and interacting protein screening to validation, our innovative portfolio of circRNA analysis will help you gain more insights and make the most impact with your research program.
     

    circRNA is a novel type of endogenous noncoding RNA that has gained attention from researchers for its involvement in multiple biological processes. It is reported that circRNA acts as miRNA sponge and RBP sponge to function in human diseases. For example, ciR-7 contains more than 70 selectively conserved miRNA target sites that can bind miR-7, facilitate a specific miR-7-AGO2 interaction, and suppress miR-7 activity in diseases including hepatocellular carcinoma, colorectal cancer and diabetes.
     

    Creative Biogene emlpoys dual luciferase reporter assay system to validate the sponge function of circRNA and detect target miRNA. This sensitive and efficient assay method is suitable for most cell types. To analysis the interaction between circRNA and RNA banding protein, Creative Biogene establishes a powerful interaction analysis system based on RNA pull-down assay and RNA immunoprecipitation (RIP) technology. Our excellent interaction analysis system can efficiently screen and validate banding protein to accelerate you research program.

    Creative Biogene is one of the leading biotechnology companies, which has the expertise and ability in providing circRNA-miRNA/RBP interaction analysis service for customers worldwide. Based on our sophisticated equipment, advanced technology and experienced staff, we will assist you with the most affordable and high-quality service to ensure your satisfaction in a timely and professional manner.

  • Oligonucleotide Immunogenicity Assessment

    IntegrateRNA's team help you perform your immunology testing. Our global team of experts partner with you to develop the best translational path for your therapeutic, provide detailed regulatory expectations, and offer technical expertise in required assay formats. By using this comprehensive approach, you'll have the most rapid and cost-effective program possible, from Proof of Concept, through safety assessment, and eventually to the clinic.

    Oligonucleotide-based therapeutics are being tested for an array of local and systemic therapeutic indications, and several compounds are approved for commercial use. As a drug class, these compounds are chemically synthesized, but have large molecular size and complex mechanism of action, and engage specific metabolic and immunologic pathways. Furthermore, given the distinct properties of the different compounds that are included in this drug class, the approach to assessing product immunogenicity should be carefully considered.

    Our Immunogenicity Expertise

    Dedicated LBA Laboratory

    • Critical reagent characterization
    • Consistent, thorough method development process
    • Performance characteristics meet/exceed regulatory requirements
    • Performance Evaluation assures successful assay validation
    • Sample analysis for pre-clinical and clinical studies

    Method Development

    • Translational sciences integration and planning
    • Customized for your molecule and mechanism of action
    • Focus on the most clinically-relevant aspects of assay
    • Optimization of format, reagents, drug tolerance, positive control
    • Preliminary cut point assessment and performance evaluation
    • Systematic transfer from development to validation

    Validation

    • Performance evaluation to assure successful validation
    • Validation Plans aligned with regulatory guidelines
    • Balanced design
    • Cut point setting
  • Oligonucleotide Linkers for Conjugation

    Despite the selectivity of oligonucleotides, they still face challenges such as short serum stability, low membrane permeability and lack of tissue selectivity. Cleavable linkers are essential in the field of oligonucleotide therapeutics as they facilitate efficient coupling of the therapeutic payload to the oligonucleotide while maintaining stability during circulation and enabling controlled release from the target site. This drug release mechanism is highly effective in oligonucleotide applications. At IntegrateRNA, we offer comprehensive services to support your oligonucleotide coupling program. We can provide different cleavable linkages to help researchers choose their oligonucleotide coupling method based on the compatibility of downstream applications.

    What We Offer?

    Suitable linkers are critical to the success of oligonucleotide therapy. Typically, the linker must remain stable in circulation and ensure safe release of the payload in the cell. Cleavable linkers are by far the most commonly used connectors in oligonucleotide design and play an important role in improving plasma stability, water solubility, target release efficiency, oligonucleotide drug distribution, and pharmacokinetics of oligonucleotide-based therapies. IntegrateRNA can provide various safe, cleavable linkers with different drug release mechanisms and stability in the circulation, including but not limited to:

    pH-Sensitive Linkers

    pH-sensitive linkers are essential in designing oligonucleotide couplers as they control the release of the oligonucleotide payload in response to pH changes. These linkers are stable in alkaline environments but highly sensitive to acidic environments, such as perylene. The low pH of endosomes (pH=5-6) and lysosomes (pH=4.8) is utilized to trigger hydrolysis of acid-unstable stilbene junctions and subsequent release of the payload. pH-sensitive linkers have several main functions, including enhanced intracellular delivery, targeted therapeutic treatment, protection from nuclease enzymes, and tunable release kinetics.

    Disulfide Linkers

    A disulfide bond consists of two sulfur atoms covalently bonded to form a disulfide bond (-SS-). The disulfide bond is relatively stable in the environment and can be doped into specific positions within the oligonucleotide sequence, allowing precise control of the affixation site, but can be cleaved by intracellular glutathione reduction to release the payload. This controlled release mechanism enhances therapeutic efficacy and minimizes off-target effects. It has multiple applications in siRNA delivery, ADCs, oligonucleotide-protein affixes, and imaging agents.

  • Lentivirus Production for Autophagy Flux Detection

    Autophagy is a highly conserved catabolic process in which long-lived cytoplasmic components or damaged organelles are sequestered by the formation of double-membraned autophagosomes. Mature autophagosomes ultimately fuse with lysosomes to form single-membraned autophagolysosomes that degrade or recycle their contents. Upon autophagy initiation, microtubule-associated protein 1 light chain 3 (LC3) is converted from LC3-I to the lipidated LC3-II and anchored to the autophagic membrane. The punctate distribution of LC3-II is regarded as a marker of autophagy induction, and it is closely related to the accumulation of autophagosomes. Lipidated LC3-II usually interacts with p62, which is a multifunctional protein that is degraded by the autophagic-lysosome pathway. The occurrence of complete autophagic flux is commonly reflected in the expression of LC3-II and p62. Currently, DNA constructs encoding fluorescent proteins fused to LC3 are widely employed for introduction into cells for monitoring autophagosome formation by fluorescence microscopy.

    QVirus™ Platform has launched series of lentiviral packaging service of autophagy related biosensors, in which GFP and/or RFP tags are fused at the C-termini of the autophagosome marker LC3, allowing to detect the intensity of autophagy flux in real-time with more accuracy, clarity and intuitiveness. These biosensors provide an enhanced dissection of the maturation of the autophagosome to the autolysosome, which capitalizes on the pH difference between the acidic autolysosome and the neutral autophagosome. Our GFP-LC3, GFP-LC3 Control Mutant, and RFP-LC3 lentiviral particles provide bright fluorescence and precise localization of LC3 to the autophagosome, enabling live cell analysis of autophagy even in difficult-to-transfect cell types.

  • CircRNA Overexpression Stable Cell Line Development

    Circular RNAs (circRNAs) are a class of single-stranded, non-coding RNAs which form a covalently closed loop. It was first reported as viroids in plants and was later detected in human cells in 1979. In a circular RNA molecule, unlike a linear RNA, the 5' and 3' ends have been joined together. The unique loop structure of circRNAs provides them with a longer half-life and more resistance to RNase R than linear RNAs. CircRNAs are produced by non-canonical splicing process which is known as "backsplicing". In the process of backsplicing, a downstream splice donor is joined to an upstream splice acceptor. According to the sequence composition of circRNAs, they are categorized into various types, such as exonic circRNA, exon-intron circRNA (EIciRNA), and circular intronic RNA (ciRNA) etc.

    Many studies have revealed that circRNAs have unique expression signatures and contribute to biological processes through a variety of functions, for example, acting as transcriptional regulators/microRNA(miRNA) sponges/protein templates, and serving as protein decoys, scaffolds and recruiters etc. The unique structure and broad expression of circRNAs makes them potential candidates for diagnostic biomarkers and therapeutic targets of a variety of diseases, such as cancers, cardiovascular diseases, neurological disorders and autoimmune diseases etc. More recently, an increasing number of investigations have been involved in the field of circRNA.

    Our Capability
    The gain-of-function and loss-of-function are approaches to investigate the roles and mechanisms of circRNAs. Cell lines stably overexpressing circRNAs are useful tools for the functional characterization of circRNAs of interest. Creative Biogene has excellent platforms for providing products and services in the field of stable cell lines and circRNAs. So far, we have developed a large number of cell lines stably overexpressing various types of genes, premade circRNA molecules and expression vectors, with more under development. Now Creative Biogene is providing a comprehensive portfolio of circRNA stable cell lines from circRNA design, vector construction, to stable cell line generation and validation. All our circRNA stable cell lines are fully validated by sequencing and real-time qPCR to make sure the sequence accuracy and high expression level.

    Highlights
    Overexpression of circRNAs in a broad type of cancer and immortal cell lines
    Stable and long-term expression of circRNAs
    Cell line models for evaluating gain-of-functions of circRNAs
    High overexpression level of circRNAs
    Good stability of circRNA overexpression
    Fully verified by QPCR and sequencing
    Mycoplasma free circRNA cell line models

  • Advancements in Microbial Genome Editing through CRISPR Plasmid Library Construction

    In recent years, the development of CRISPR-Cas technology has revolutionized the field of molecular biology, allowing for precise and efficient genome editing in a variety of organisms. One of the key applications of CRISPR-Cas technology is the construction of CRISPR plasmid libraries, which can be used to systematically target and modify genes within microbial genomes.

    Microbial genome CRISPR plasmid libraries are powerful tools that enable researchers to study gene function, pathway dynamics, and genetic interactions on a large scale. These libraries consist of a collection of plasmids, each containing a unique guide RNA (gRNA) sequence designed to target a specific gene or genetic sequence within the microbial genome. By delivering these plasmids into bacterial cells, researchers can induce targeted mutations, knockouts, or gene insertions at precise genomic loci.

    The construction of a microbial genome CRISPR plasmid library involves several key steps. First, a library of gRNA sequences is designed using bioinformatics tools to target genes of interest within the microbial genome. These gRNA sequences are then synthesized and cloned into a plasmid vector containing the Cas9 nuclease gene. The resulting plasmid library is transformed into bacterial cells, creating a diverse pool of genetically modified strains that can be screened and analyzed for specific phenotypes.

    One of the major advantages of microbial genome CRISPR plasmid libraries is their scalability and flexibility. Researchers can easily expand and customize the library by adding new gRNA sequences or targeting different genes of interest. This flexibility allows for the rapid and high-throughput screening of gene function and genetic interactions in microbial systems.

    Furthermore, microbial genome CRISPR plasmid libraries have been successfully employed in a wide range of studies, including functional genomics, drug discovery, and biotechnology applications. By leveraging the power of CRISPR-Cas technology, researchers can accelerate their understanding of microbial physiology and develop novel strategies for bioengineering and bioproduction.

    In conclusion, the construction of microbial genome CRISPR plasmid libraries represents a cutting-edge approach for precise genomic editing in bacteria. These libraries provide a valuable resource for studying gene function and genetic interactions at a genome-wide scale, enabling researchers to unlock the full potential of microbial systems for biotechnological applications.
     
  • Custom Synthesized sgRNA: A Versatile Tool for Precision Gene Editing in Biological Research

    In the field of biological research, the ability to precisely edit the genetic material of organisms has revolutionized the way scientists study and understand biological processes. One of the most powerful tools for achieving this precision is the use of synthetic guide RNA (sgRNA), a key component of the CRISPR/Cas9 genome editing system. Custom synthesized sgRNA offers researchers a flexible and efficient means of targeting specific genetic sequences for modification, opening up a world of possibilities for genetic manipulation and experimentation.

    The beauty of sgRNA lies in its specificity and versatility. By designing a custom sequence of sgRNA that complements a target DNA sequence, researchers can guide the Cas9 enzyme to precisely cut and edit that portion of the genome. This targeted editing allows for the introduction of mutations, insertions, or deletions at precise locations within the genome, enabling researchers to investigate the function of specific genes or correct genetic defects.

    Custom synthesized sgRNA can be tailored to target virtually any gene of interest, making it an invaluable tool for a wide range of biological research applications. Whether studying gene function in model organisms, developing disease models, or investigating the mechanisms of genetic disorders, sgRNA can be customized to suit the specific research needs of the investigator. Additionally, the ease of design and synthesis of sgRNA allows for rapid and cost-effective experimentation, reducing the time and resources required for genetic manipulation.

    Furthermore, the development of custom synthesized sgRNA has expanded the possibilities of genome editing beyond just model organisms. Researchers are now using this technology to engineer crops for improved yield and resistance to pests, develop gene therapies for genetic diseases in humans, and even create novel biotechnological applications. The potential for custom synthesized sgRNA to revolutionize fields such as agriculture, medicine, and biotechnology is immense, driving innovation and discovery in a wide range of scientific disciplines.

    In conclusion, custom synthesized sgRNA is a powerful tool that has transformed the field of biological research by enabling precise and targeted genetic editing. Its versatility and ease of customization make it an essential component of the researcher's toolkit, facilitating advances in genetics, molecular biology, and biotechnology. As scientists continue to harness the capabilities of sgRNA in their experiments, the potential for groundbreaking discoveries and transformative applications in the life sciences is limitless.