Creative Biolabs was founded by an experienced team of proven biotech executives to develop oncology research tools that can potentially eradicate intractable tumors. we are now developing a proprietary whole-organism combinatorial and cumulative genome editing platform for precision tumor lineage tracing. With this approach, we can get an accurate, comprehensive lineage tree of tumor cells.

Introduction to Tumor Lineage Tracing Platform Development

Over the past few decades, Creative Biolabs has developed a diverse range of innovative tumor lineage tracing methods to establish lineage information for oncogenesis, tumor progression, or tumor recurrence. Generally, a series of tumor cellular labeling systems such as dyes and enzymes, recombinant enzyme-mediated reporter genes, foreign DNA incorporation, and naturally occurring somatic mutations have been successfully validated and widely used in our labs.

However, existing techniques still possess certain limitations when it comes to reconstructing cell lineages in large-scale multicellular systems. For instance, in our dye and reporter gene-mediated systems, lineage relationships between offspring cells can not be identified. Moreover, distinguishing numerous branches resulting from independent and equal labeling of two or more tumor cells is still challenging.

Fig.1 Decision Tree for Choosing an Appropriate Lineage Tracing Strategy.¹

Although different DNA barcodes or somatic mutations can be introduced to identify lineages and sublineages within a single organism, these systems still require expensive whole-genome sequencing analysis which is not easily compatible with in situ readouts for small numbers of tumor cell samples.

Therefore, we are now dedicated to designing a comprehensive lineage-tracing system for multiple tumor cells. Our system enables precise and incremental labeling of tumor cells and their offspring. Each tumor cell will be labeled by distinct markers that can be utilized to construct lineage trees. Also, a single tumor cell can provide a complete set of labeled readouts.

Whole-organism Combinatorial and Cumulative Genome Editing Lineage Tracing System

In Creative Biolabs, our whole-organism combinatorial and cumulative genome editing system has been considered as a lineage tracking approach for new genome editing techniques based on multiple synthetic targets, where we consolidate the diversity of various sequence combinations into a concise, multi-target barcode that contains abundant information. The gene-edited barcodes effectively facilitate the retrieval of multiple single-cell tumor information from each sequencing readout.

Fig. 2 Reconstruction of a Synthetic Lineage Based on Genome Editing And Targeted Sequencing of Edited Barcodes.²

Currently, we have synthesized a continuous sequence of 8 gene-editing targets, separated by 3-5 base pair (bp) linkers, to combine and cumulatively edit a unique genomic barcode combination element. Among them, the first target perfectly matched a single guide RNA (sgRNA), while the remaining off-target sites of the sgRNA are ranked from most active to least active. Secondly, we transfected the same editing reagent into several separate batches of cultures to evaluate the repeatability of this system. In our labs, this unique barcode element can edit multiple target sites, resulting in a combinatorial diversity of alleles. With single sequencing, we can further read allelic DNA or RNA. In addition, the rates and patterns of these barcodes can be adjusted by using targets with different active and/or off-target sequences. The recombination target can also be iterated by designing new barcodes, adjusting the regents' concentration, and editing the delivery patterns.

Why Choose Us?

• One-stop tumor lineage tracing solution
• Cost-saving discovery strategy
• Mature whole-organism combinatorial and cumulative genome editing platform
• Fast turnaround time

We, at Creative Biolabs, have a deep legacy of scientific innovation. As pioneers in the field of human oncology imaging, we believe that science and innovation can make a difference for humankind. From the very beginning, our focus has always been on exploiting a multifunctional whole-organism combinatorial and cumulative genome editing system to help address cancer cell fates. Some of our scientists, including our co-founders, have been conducting fundamental research for decades to understand lineage tracking and its mechanisms to improve our technologies. If you are interested in our services, please do not hesitate to contact us for more details.

References

1. Kretzschmar, K.; et al. Lineage tracing. Cell, 2012, 148(1): 33-45.
2. McKenna, A.; et al. Whole-organism lineage tracing by combinatorial and cumulative genome editing. Science, 2016, 353(6298): aaf7907.

For Research Use Only | Not For Clinical Use