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. 2020 Nov 12;16(1):198–211. doi: 10.1016/j.stemcr.2020.10.007

Figure 3.

Figure 3

Viral RNA of LVs Showed Length-Dependent Decrease in Completeness and Coverage in RNA Sequencing

(A) Comparison of titer versus vector length (bars represent mean with SEM; n = 3 independent experiments; Pearson correlation, r = −0.86, p < 0.01).

(B) Complete RNA versus vector length in a panel of β-globin vectors of different lengths (bars represent mean with SEM; n = 3 independent experiments; Pearson correlation exclude Globe-AS3, r = −0.67, p < 0.05). All β-globin vectors were packaged and assayed concurrently. Viral RNA was extracted from vector particles.

(C) Percentage of maximum reads of EFS-ADA, Globe-AS3-FB, and Lenti/βAS3-FB versus vector length (n = 3 independent experiments). Schematic representation of the viral RNA is displayed below. LVs were packaged in three independent experiments. Total RNA was extracted from 140 μL unconcentrated viral supernatant followed by DNase treatment. RNA was fragmented to an average of 400 bp, reverse transcribed by random priming, and sequencing adaptors ligated. The library was sequenced on an Illumina HiSeq 3000.