An exogenous encoding sequence based on DNA data storage technology and its application in assisted reproductive technology
Description
Safety and ethical issues are the primary concerns for assisted reproductive technology (ART). However, confusion and contamination of samples are common problems in embryo laboratories, preimplantation genetic test (PGT) laboratories, and third-party medical testing laboratories due to large sample numbers and complex procedures. Once these problems occur, they are often difficult to trace, posing risks and ethical challenges to hospital reproductive centers, third-party medical testing laboratories, and patient families. Therefore, it is necessary to establish an effective and feasible tracing system to ensure sample safety. In this study, we designed an exogenous encoding sequence (EES) based on DNA data storage technology, which provide a unique identification code for each in vitro cultured embryo, effectively avoiding potential risks and ethical problems caused by sample confusion and contamination. This exogenous encoding sequence is a DNA molecule that is non-toxic and structurally stable. We verified that a small amount of exogenous encoding sequence (6*109 copies/uL) can be amplified together with embryo biopsy cells and detected by various sequencing methods without affecting copy number variants (CNVs). Furthermore, if there is contamination from other samples at a proportion of more than 5%, it can also be identified through the encoding information of the exogenous encoding sequence. Our study proves that the exogenous encoding sequence designed based on DNA data storage technology is effective and reliable, and can be applied in hospital reproductive centers and third-party medical testing laboratories to improve the safety of in vitro cultured embryos and avoid potential ethical problems in the future.
Files
Steps to reproduce
The obtained cells undergo whole genome amplification (WGA) and NGS library preparation using the ChromInstTM Universal Library Preparation Kit (Cat No.: XK-008-24/XK-008-96, Manufacturer: Xukang Medical Technology (Suzhou) Co., Ltd) following the provided operational guidelines. Initially, after cellular lysis, the liberated genomic DNA is primed with a random primer library, initiating pre-amplification which is succeeded by exponential amplification to achieve a 2 µg output. Subsequently, the ChromInstTM Universal Library Preparation Kit is employed for WGA and library preparation. Validation of the NGS library's quality is carried out through Qubit 3.0 and 1.5% agarose gel electrophoresis. Sequencing is conducted on the Illumina platform, yielding approximately 2 million sequencing reads per library.