Crispr barcoding: method and kit for labeling and detecting a population of endonuclease-treated cells

The invention relates to methods, compositions and kits for labelling and detecting endonuclease-treated cells, and most preferably eukaryotic cells. The CRISPR-barcoding system could be used as an alternative tool to the classical lentiviral DNA barcode libraries, ensuring the detection of thousands of distinct barcodes through qPCR or deep-sequencing. This barcoding approach enables tracing of the mutated cells immediately after DNA editing without the need to derive clones, thus providing a unique means to investigate the effects of different kinds of genomic modifications, regardless of their potential impact on cell growth, in a broad range of functional assays. This strategy represents a high-resolution tracking of single specific cancer cells allowing to identify even rare pre-existing resistant subclones potentially involved in mechanisms of acquired resistance to therapy. The main proofs of concept reported are related to cancer models, nevertheless this technology can be implemented in different fields of biological research.

Keywords: CRISPR/Cas9, barcoding, Gene editing
Patent Application number: EP 15 306 698.0
WO2017068120A1
Publications:
CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and Functional Analysis of Oncogenic Driver Mutations. Mol Cell. 2016 Aug 4;63(3):526-38. doi: 10.1016/j.molcel.2016.06.017. Epub 2016 Jul 21. Modeling intratumor heterogeneity through CRISPR-barcodes. Mol Cell Oncol. 2016; 3(6): e1227894. CRISPR/Cas9 editing of the genome for cancer modeling. Methods. 2017 May 15;121-122:130-137. doi: 10.1016/j.ymeth.2017.03.007. Epub 2017 Mar 10.

Reference:

BIO14199-R1

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Patent filling date: 22-10-2015

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