CRISPR/Cas9-deaminase enables robust base editing in Rhodobacter sphaeroides 2.4.1.

ABSTRACT:

Background

CRISPR/Cas9 systems have been repurposed as canonical genome editing tools in a variety of species, but no application for the model strain Rhodobacter sphaeroides 2.4.1 was unveiled.

Results

Here we showed two kinds of programmable base editing systems, cytosine base editors (CBEs) and adenine base editors (ABEs), generated by fusing endonuclease Cas9 variant to cytosine deaminase PmCDA1 or heterodimer adenine deaminase TadA-TadA*, respectively. Using CBEs, we were able to obtain C-to-T mutation of single and double targets following the first induction step, with the efficiency of up to 97% and 43%; while the second induction step was needed in the case of triple target, with the screening rate of 47%. Using ABEs, we were only able to gain A-to-G mutation of sin

SUBMITTER: Luo Y 

PROVIDER: S-EPMC7183636 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature