Vectorology - GEG Tech top picks

In this work, the scientists describe adaptations to the MGS-PCR protocol for the IlluminaMiSeq platform for gammaretroviral, lentiviral and foamy virus vectors. Using unique combinations of indices in the PCR primers, up to twelve samples can be pooled into one sequencing reaction to reduce costs.  Modified genomic sequencing PCR using the MiSeq platfom led to paired-end reads of average length 265 bp. These longer sequence reads allow for improved localization of retroviral vector integration sites to published genomes.

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The scientists have developed a rapid and inexpensive method for measuring clone size based on random shearing of genomic DNA, minimal exponential PCR amplification, and shear site counts as a quantitative endpoint. Samples were analyzed from transplanted pigtail macaques and from a participant in our X-linked severe combined immunodeficiency (XSCID) lentiviral vector trial and yielded controlled and quantitative results in all cases.One case of early clonal dominance was detected in a monkey transplanted with limiting numbers of transduced HSCs while the clinical samples from the XSCID trial participant showed highly diverse clonal representation.

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