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ABE8e with Polycistronic tRNA-gRNA Expression Cassette Sig-Nificantly Improves Adenine Base Editing Efficiency in Nicotiana benthamiana .
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- معلومة اضافية
- المصدر:
Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE
- بيانات النشر:
Original Publication: Basel, Switzerland : MDPI, [2000-
- الموضوع:
- نبذة مختصرة :
Adenine base editor containing TadA8e (ABE8e) has been reported in rice. However, the application of ABE8e in other plant species has not been described, and the comparison between ABE8e and ABE7.10, which is widely used in plants, has also been poorly studied. Here, we developed the ABE8e with the polycistronic tRNA-gRNA expression cassette (PTG-ABE8e) and PTG-ABE7.10 and compared their A-to-G editing efficiencies using both transient and stable transformation in the allotetraploid Nicotiana benthamiana . We found that the editing efficiency of PTG-ABE8e was significantly higher than that of PTG-ABE7.10, indicating that ABE8e was more efficient for A-to-G conversion in N. benthamiana . We further optimized the ABE8e editing efficiency by changing the sgRNA expression cassette and demonstrated that both PTG and single transcript unit (STU) enhanced ABE8e efficiency for A-to-G conversion in N. benthamiana . We also estimated the potential off-target effect of PTG-ABE8e at potential off-targeting sites predicted using an online tool in transgenic plants, and no off-target editing event was found for potential off-targeting sites selected, indicating that ABE8e could specifically facilitate A-to-G conversion. Our results showed that ABE8e with PTG structure was more suitable for A-to-G conversion in N. benthamiana and provided valuable clues for optimizing ABE tools in other plants.
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- Grant Information:
XDA24030404 Chinese Academy of Sciences Strategic Leading Science and Technology Project; 2019YFD1000201 National Key R&D Program of China
- Contributed Indexing:
Keywords: ABE; ABE8e; CRISPR; PTG; STU; base editor
- الرقم المعرف:
0 (RNA, Guide, CRISPR-Cas Systems)
- الموضوع:
Date Created: 20210602 Date Completed: 20210617 Latest Revision: 20240104
- الموضوع:
20240104
- الرقم المعرف:
PMC8198424
- الرقم المعرف:
10.3390/ijms22115663
- الرقم المعرف:
34073486
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