Modular AAV-based base editors

The CBE plasmids including pCAG-CBE4max-SpG-P2A-EGFP [52 (link)] (Addgene #139,998), pCMV_AncBE4max [23 (link)] (Addgene #112,094) and pAAV-pCMV-dSaCas9-VP64-pU6-sgRNA (Addgene #158,990) were obtained from Addgene (Watertown, MA). All gRNA oligos were annealed and cloned into pLenti-ogRNA via BsmBI site. The hAAT promoter sequence was PCR amplified from the genomic DNA of HEK293 cells. The hAAT promoter, Gp41-1 intein split base editor fragments, and the human U6-driven gRNA fragment, were PCR amplified and inserted into the AAV transfer plasmids pZC0031 and pZC0033 as described previously [22 (link)].

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Zuo Y., Zhang C., Zhou Y., Li H., Xiao W., Herzog R.W., Xu J., Zhang J., Chen Y.E, & Han R. (2023). Liver-specific in vivo base editing of Angptl3 via AAV delivery efficiently lowers blood lipid levels in mice. Cell & Bioscience, 13, 109.

Publication 2023

pCMV_AncBE4max

Genomic Hek293 cells Human Intein Oligos Plasmids

Corresponding Organization :

Other organizations : Indiana University – Purdue University Indianapolis, University of Michigan–Ann Arbor, Michigan Center for Translational Pathology, The Ohio State University

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Variable analysis

independent variables

CBE plasmids including pCAG-CBE4max-SpG-P2A-EGFP, pCMV_AncBE4max, and pAAV-pCMV-dSaCas9-VP64-pU6-sgRNA
GRNA oligos cloned into pLenti-ogRNA
HAAT promoter sequence PCR amplified from HEK293 cells
Gp41-1 intein split base editor fragments
Human U6-driven gRNA fragment

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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