<b>Engineered </b><b>VPg</b><b> </b><b>saRNA achieves cap-independent,</b><b> </b><b>low-immunogenic and precise</b><b> </b><b>encoding of therapeutic proteins </b><b>in vivo</b><b>.</b>
<p dir="ltr">AsPC-1 pancreatic cancer cells were transfected with the VPg-GSDMDeng saRNA vaccine and subsequently cultured for 15 consecutive passages to evaluate sequence stability during prolonged intracellular replication. Total RNA was isolated from both the first-generation (P1)...
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2025
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| 总结: | <p dir="ltr">AsPC-1 pancreatic cancer cells were transfected with the VPg-GSDMDeng saRNA vaccine and subsequently cultured for 15 consecutive passages to evaluate sequence stability during prolonged intracellular replication. Total RNA was isolated from both the first-generation (P1) transfected cells—used as the non-mutated reference control—and the 15th-generation (P15) cells. High-fidelity reverse transcription PCR (RT-PCR) was performed using a proofreading DNA polymerase to minimize polymerase-induced errors. The resulting amplicons were subjected to bidirectional Sanger sequencing using multiple forward primers (M13F, seq1F, seq2F, seq3F) to ensure complete coverage of the coding region and to detect low-frequency variants.</p><p dir="ltr">All raw chromatogram files (.ab1) and derivative sequence files (.seq) have been deposited in the public repository. Each .ab1 file corresponds to an individual sequencing reaction and retains unprocessed fluorescence trace data, peak calls, and Phred quality values.</p> |
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