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rLuc Lentiviral Expressed Libraries for In Vivo Bioluminescent Imaging (BLI)

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Cellecta CloneTracker XP-rLuc have the same design as the its other CloneTracker XP Libraries, with a barcode interested 3′-UTR of the reporter transcript so it is detectable by RNA sequencing. Rather than a fluorescent protein marker, however, CloneTracker XP-rLuc library have a red-shifted luciferase (rLuc) derived from Photinus pyralis (Ppy RE9 mutant) as a reporter for bioluminescent imaging (BLI).
Luciferase reporters provide a useful tool for indirect cell labelling and tracking of cell fates in vivo. While the BLI technique has been successfully used for monitoring of migration, growth, and viability associated with drug treatment of transplanted cancer, stem, or immune cells in mouse models, standard luciferase assays in animal models are particularly sensitive because they are not subject to high background as a result of tissue auto-fluorescence. The codon-optimized Photinus pyralis far-red luciferase in the XP-rLuc Library offers enhanced sensitivity and resolution in deep tissue-50-100-fold better light intensity than the click beetle (Renilla) enzyme.
Cellecta pScribe6-rLuc-CloneTracker-XP-Barcode-Vector-Map

Depending on the construction of the library vector, a clonal barcode construct can be constructed so that that the barcode sequence can not only be read from genomic DNA but also expressed so it is detectable by sequencing the RNA cell fraction. For example, Cellecta offers barcode libraries (i.e., CloneTracker XP™ Libraries) where the barcode is located in the 3’-UTR of the antibiotic selection gene for the vector, as shown in the figure.  With this configuration, the clonal barcodes can be detected in single cell bead-based RNA expression profiling assays, such as 10X Genomics System, BD Rhapsody, or Dolomite Bio’s Nadia system, so that changes in gene activation in different clonal populations can be assayed over the course of an experiment to identify sub-populations of progeny with advantageous or harmful phenotypes relative to the specific conditions, such as increased drug resistance or sensitivity.

The CloneTracker XP-rLuc Library can be transduced into cells to create a barcoded cell population which, when engrafted into mice, can be used for monitoring tumor formation and metastasis growth. BLI of rLuc-tagged cells in vivorequires the use of an appropriate imager for detecting photons generated from D-Luciferin substrate by CCD cameras able to sample the entire visible and near infrared spectrum. For details on in vivo imaging, please refer to the standard protocols provided with your instrument, for example, In vivo Bioluminescence Imaging of Luciferase-labeled Cancer Cells protocol for BLIusing the IVIS Spectrum imager from PerkinElmer.
Mouse Bioluminescent imaging Cellecta Clonetracker XP rLuc

 More information on Cellecta’s CloneTracker XP-rLuc 10M-3' Lentiviral Barcode library offering available here.