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Figure 1

From: In vivo quantification of formulated and chemically modified small interfering RNA by heating-in-Triton quantitative reverse transcription polymerase chain reaction (HIT qRT-PCR)

Figure 1

Efficient small interfering RNA (siRNA) quantification requires release from the formulation and strand separation. Amplification curves of unmodified (antisense (AS)), modified AS (mod AS), duplex (Duplex), modified duplex (mod Duplex), and lipid nanoparticle (LNP)-KC2-formulated modified duplex (KC2 mod Dup) siRNAs. siRNAs were diluted as indicated either in water or in 0.25% Triton. Then were treated as indicated either at room temperature (25°C) or heated to 95°C for 10 min and added directly into reverse transcription reactions: (a) water; 25°C, (b) water, 95°C, (c) water, 95°C for mod AS, mod Dup and for KC2-mod Dup, both in 25°C and 95°C, and (d) 0.25% Triton-phosphate-buffered saline (PBS); 95°C. The linear regression of the amplification curve for chemically modified AS siRNA is shown (inear fit (mod AS)). The average cycle threshold (AvCt) values (n = 3) indicate signals in single PCR reactions containing siRNA equivalent concentrations ranging between 3.35 × 10-2 to 3.35 × 10-7 pmol.

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ISSN: 1758-907X