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Abstract:
TALEN-mediated plant genome editing has the advantages of high specificity, independence from epigenetic modification, lack of PAM limitation, and feasibility for organelle editing due to no need for RNA binding. However, the assembly of TALEN vectors was difficult and laborious, affecting the popularity of TALEN genome editing in plants. In this study, a novel TALEN-based plant genome editing tool, ZQTALEN, was developed. This system comprises a total of nine plasmids categorized into three types. The TALE repeat units are obtained through PCR utilizing the template vector as the amplification template. These units are then assembled sequentially: first into donor vectors to form entry vectors, and subsequently transferred to destination vectors, resulting in the final binary vector. Characterized by the optimization of codon usage, assembly method of TALE repeat array, and vector backbone components, ZQTALEN has the advantages of easy, flexible and efficient assembly and less repeated sequences in the final vector. Using the ZQTALEN system, a TALEN binary vector was successfully constructed to target the endogenous Nramp5 gene in rice, resulting in the high-frequency acquisition of rice mutants. The ZQTALEN system has provided a versatile tool for genetic research in plants.
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PLANT MOLECULAR BIOLOGY
ISSN: 0167-4412
Year: 2025
Issue: 1
Volume: 115
3 . 9 0 0
JCR@2023
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ESI Highly Cited Papers on the List: 0 Unfold All
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30 Days PV: 0
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