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For researchers investigating tumor progression, drug resistance, differentiation, development, hematopoiesis, and other related areas, there is considerable interest in understanding the heterogeneity of cell populations in both cultured and in vivo settings, by incorporating stable, heritable, and sequenceable barcodes in individual cells or cell populations.
Label individual cells in a population with stable, genomically integrated, single, identifiable, sequenceable barcodes for cell tracking and analysis of biological variations across populations. Library constructs integrate into the genomic DNA after transduction and can be detected by genomic DNA amplification.
With the CloneTracker XP™ libraries, the barcodes can also be detected in the cDNA generated when running single-cell RNA sequencing procedures.
Sub-libraries with a unique set of barcodes are also available to label several cell populations with distinct non-overlapping barcodes.
Libraries of millions of barcodes encapsulated in VSV-g pseudotyped lentiviral particles efficiently transduce and integrate into the host cell genome of virtually any mammalian cells. When a cell population is transduced at a low number of viral particles to cells (low MOI), individual cells pick up and incorporate a single barcode into their genomic DNA. As barcoded cells divide, the barcode sequences will be passed onto the cell progeny. Cells can be treated, grown for several passages, frozen and thawed, and the sequences within the lentiviral vector will remain in the host cell. After selection, treatment, or differentiation, just extract genomic DNA, amplify, and sequence to identify and quantify barcodes present in the selected cell population.
Cellecta’s CloneTracker XP Libraries have been designed with the barcodes located at the 3′ end of the selection transcript. As a result, the barcodes are transcribed during poly-A or switch-oligo (aka, SMART oligo) cDNA synthesis so they are expressed and quantifiable using RNA-Seq.
Single-cell RNA-Seq analysis is an important tool to analyze individual cell responses. However, important genes and pathways may not be active in all cells. Certain responses may only occur in a portion of cells in a population, leading to divergent phenotypes when selected in some manner, for example, during drug treatment. One way to detect a small number of cells with a biologically interesting phenotype is to label individual cells with uniquely identifiable barcodes. Responses of each of the clonal progeny from this initial barcoded population can be assessed, and data correlating to how experimental conditions, differentiation, or developmental processes affect distinct groups of cells in a population can be investigated.
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