Abstract
The piggyBac (PB) transposon is the most widely used vector for generating transgenic silkworms. The stability of the PB transposon in the receptor is a serious concern that requires attention because of biosafety concerns. In this study, we found that the transgene silkworm developed loss of reporter gene traits. To further investigate the regularity, we traced the genes and traits of this silkworm. After successful alteration of the silkworm genome with the MASP1 gene (named red-eyed silkworm; RES), silkworm individuals with lost reporter genes were found after long-term transgenerational breeding and were designated as the white-eyed silkworm (WES). PCR amplification indicated that exogenous genes had been lost in the WES. Testing was conducted on the PB transposons, and the left arm (L arm) did not exist; however, the right arm (R arm) was preserved. Amino acid analysis showed that the amino acid content of the WES changed versus the common silkworm and RES. These results indicate that the migration of PB transposons in Bombyx mori does occur and is unpredictable. This is because the silkworm genome contains multiple PB-like sequences that might influence the genetic stability of transgenic lines. When using PB transposons as a transgene vector, it is necessary to fully evaluate and take necessary measures to prevent its re-migration in the recipient organism. Further experiments are needed if we want to clarify the regularity of the retransposition phenomenon and the direct and clear association with similar sequences of transposons.
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Funding
This work was supported by the National Natural Science Foundation of China (No. 31830094), Chongqing postgraduate research innovation project (Grant No. CYS19110) and funds from the China Agriculture Research System (No. CARS-18-ZJ0102).
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XJ conducted molecular lab work, data analysis, sequence alignments, participated in the design of the study, and drafted the manuscript; XP, YY and QG conducted statistical analyses and critically revised the manuscript; TZ, ML, GZ, and FD developed the study design and helped draft the manuscript. All of the authors gave final approval for publication and agree to be held accountable for the work performed therein.
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All animal studies were conducted in compliance with the protocols approved by the National Centre of Animal Science Experimental Teaching of Southwest University of China.
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Jia, X., Pang, X., Yuan, Y. et al. Unpredictable recombination of PB transposon in Silkworm: a potential risk. Mol Genet Genomics 296, 271–277 (2021). https://doi.org/10.1007/s00438-020-01743-0
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DOI: https://doi.org/10.1007/s00438-020-01743-0