Millions of Defined Sequenceable Barcodes for Clonal Cell Tracking

The ability to identify and quantify the size of clonal cell populations produced by each cell in a founder population offers researchers a powerful tool for understanding how groups cells grow and proliferate in all sorts of conditions. This sort of clonal cell tracking enables cell biologists, immunologists, cancer researchers, for example, to analyze how large cell populations proliferate in various environments, progenitor cells differentiate and acquire new phenotypes, and tumor masses evolve.

Most cell tracking experiments have used fluorescence, bioluminescence, or immunofluorescence labeling which, while effective, only enables effective tracking of a handful of cells since the colors or spectra used are limited. Also, tracking in certain models, such as in vivo systems can be complicated by the direct visualization of the labeled cells required using these dye-based approaches. In general, these approaches are useful to track a pool of cells but only provide limited information about the size, distribution, or localization of the distinct progeny produced by each cell in large or complex population.

Recently, Cellecta released the CellTracker™ Lentiviral Barcode Library that provides an alternative tracking tool to fluorescent or colorimetric dyes. This library consists of lentiviral plasmid constructs, each labeled with a unique nucleotide barcode that can be easily identified by DNA sequencing. There are millions of uniquely identifiable defined barcodes in the library due to the unique way the library was made. Instead of randomly generating sequences, that nucleotide barcodes were specifically designed, synthesized, and cloned so as to to produce a library with a distinct set of well-defined sequences optimized for HT sequencing.

As a result, each individual cell in a complex population of millions can be genetically tagged with a unique DNA sequence. Since this is done using lentiviral transduction, these sequenceable genetic tags are stably integrated into the genomic DNA of the cells, and so, become a permanent heritable marker. The genetic sequence is passed on with the genomic DNA is replicated so all the progeny of the original cell that was transduced in the founder population contain the same barcode.  At any point, then, it is possible to assess, in decedent populations, the sizes of each clonal population derived from the originally transduced cells by just harvesting the cells and sequencing the genetic tags in their genomic DNA.

Cellecta Clonal Cell Tracking Barcode Library construction and quality control diagram


Leave a comment

Comments will be approved before showing up.


Also in Cellecta Blog & News

Inducible Cas9 Expression in a Single Lentiviral Vector

Introducing Inducible Cas9 Expression in a Single Lentiviral Vector to make cells capable of high Cas9 expression for a limited time during which CRISPR-mediated targeted rearrangements can occur, and then shut off Cas9 expression for downstream assays with the modified cells.
Read More
Insertion of 10X Genomics' Capture Sequences Does Not Affect HEAT-Tracr sgRNA Efficacy

Perturb-Seq or CROP-Seq screens make use of single-cell RNA-Sequencing in conjunction with a pooled CRISPR library to identify transcriptional changes and, by implication, activation or deactivation of cellular pathways related to phenotypic changes produced by specific sgRNA-mediated gene knockouts.
Read More
Core Population of Cancer Stem Cells Mediates Therapeutic Resistance in Tumors

Researchers at MD Anderson Cancer Center recently used a Cellecta CloneTracker Barcode Library to label patient-derived xenograft (PDX) cells and establish a stable population of aggressive tumorigenic cells with a specific set of barcodes. With this population of barcoded tumorigenic clones, the investigators...
Read More