Creative Biolabs is a biotechnology company focused on the research and development of tumor-based immunotherapies for cancer diagnosis. With more than a decade of research, our versatile team of scientists is going deeper into creating unmatched customized Sleeping Beauty transposon system barcode lineage tracing services to reveal the fate of individual tumor cells.

Introduction to Sleeping Beauty Transposon System

In Creative Biolabs, the Sleeping Beauty transfection vector system has become an efficient tool to insert exogenous DNA into the host cell genome. The system is very simple and requires only plasmid transfection (no viral transduction) to permanently insert the target gene you are interested in into the host cell genome.

In general, our Sleeping Beauty transposition vector system consists of two E. coli vectors. The first plasmid is used as a helper vector that encodes a transposable enzyme. The other plasmid is a transposon vector containing a transposon region surrounded by two reverse/forward repeats (IR/DR). Genes that normally need to be delivered to the host cell will be cloned into this region. Up to now, our Sleeping Beauty transposon system and its helper vector have been optimized for high copy number replication and can efficiently transfect a wide range of target cells with high levels of expression.

Advantages

• Permanent DNA integration: this system can realize transposon permanent integration in the host cell genome DNA.
• Rapid and simple: the transfection process does not require virus packaging.
• High performance: no immunogenicity and no special requirement for carrying sequence.
• Targeted Focus: multiple tumor-specific antigens.

Fig.1 Pipeline for Selecting Candidate Cancer Genes From SB Screens for Further Study.¹

Sleeping Beauty Transposon System Barcode Lineage Tracing Platform

Currently, our scientists are dedicated to developing a novel tumor lineage tracing technology based on the Sleeping Beauty transposon system and DNA barcode technology. Our platform makes it possible to develop new oncology research tools and improve existing lineage tracing methods.

To track cells by unique barcodes in complex tumor models, we continuously optimize barcoding synthesis techniques. For example, we can randomly mutate a certain length of DNA base, or base pair, to produce a series of sequences of DNA barcodes in each cell. Moreover, Sleeping Beauty-mediated insertion and deletion mutations provide more options for barcode generation in our labs.

From in vitro tumor cell models to animal models, our lineage tracking platform fills the knowledge gap in most aspects of tumor progression. In a recent project, the Sleeping Beauty transposon lineage tracking technology combined with advanced methods such as genetic barcoding and sequencing technology has given us a more accurate understanding of the origin and development of cancers. Our platform is capable of tracking the occurrence, proliferation, and recurrence of individual tumor cells. In addition, Our lineage analysis services have also expanded to tumor modeling, drug screening, cell plasticity research, as well as personalized drug discovery.

Fig. 2 Lineage Tracing With the Sleeping Beauty (SB) Mouse Model.²

The procedure of in vitro barcode assay:

(1) Isolation of tumor cell populations of interest;

(2) Barcode production by the Sleeping Beauty transposon system;

(3) Transplanting or cultivating bar-labeled cells to develop into mature lineages;

(4) Isolation of mature cell population;

(5) PCR barcode expansion;

(6) Next-generation barcode sequencing;

(7) Barcode data processing and analysis;

Customize Sleeping Beauty Transposon Barcoding Mouse Models

Normally, our Sleeping Beauty transposase-based barcoding mouse models consist of three elements:

1. A reverse tetracycline-controlled transactivator element
2. A Sleeping Beauty inducible expression element
3. A single cognate transposable Tn element

Benefits

• The Right Combination: Our expertise in leading-edge lineage tracing and Sleeping Beauty transposase-based barcoding mechanisms gave rise to the discovery of a new approach to human oncology research.
• Building on Success: After witnessing the potential of lineage tracing technology to reveal the fate of tumor cells, we used this mechanism to prevent the occurrence and recurrence of tumors.
• A Well-Mature and Customer-Driven Approach: At Creative Biolabs, we are unwavering in our commitment to expanding our Sleeping Beauty transposon system barcode lineage tracing platform to provide data support for the treatment and prevention of tumors.

Creative Biolabs is at the forefront of developing a new human oncology imaging strategy by harnessing its flexible Sleeping Beauty transposon system barcode lineage tracing technology. By delivering multiple barcodes, we can accurately detect changes in multiple generations of tumor cells. We apply our technology to designing off-the-shelf, most efficient, and scalable tumor lineage tracing systems. If you are interested in our services, please contact us for more details. Let us know what you need and we will accommodate you. We look forward to working with you in the future.

References

1. Tschida, B. R.; et al. Mouse models of cancer: Sleeping Beauty transposons for insertional mutagenesis screens and reverse genetic studies. In Seminars in cell & developmental biology, 2014, 27: 86-95.
2. Carrelha, J.; et al. Single-cell lineage tracing approaches in hematology research: technical considerations. Experimental hematology, 2020, 89: 26-36.

For Research Use Only | Not For Clinical Use