Cytosine base editing systems with minimized off-target effect and molecular size
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Abstract Structure-based rational engineering of the cytosine base editing system Target-AID was performed to minimize its off-target effect and molecular size. By intensive and careful truncation, DNA-binding domain of its deaminase PmCDA1 was eliminated and additional mutations were introduced to restore enzyme function. The resulting tCDA1EQ was effective in N-terminal fusion (AID-2S) or inlaid architecture (AID-3S) with Cas9, showing minimized gRNA-independent off-targets, as assessed in yeast and human cells. Combining with the smaller Cas9 ortholog system, the smallest cytosine base editing system was created that is within the size limit of AAV vector.
2006 ◽
Vol 26
(6)
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pp. 2297-2308
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1999 ◽
Vol 96
(9/10)
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pp. 1580-1584
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1985 ◽
Vol 260
(4)
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pp. 2301-2306
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1982 ◽
Vol 257
(11)
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pp. 6102-6105