BibTex format

author = {Wu, Y and Li, Y and Liu, Y and Xiu, X and Liu, J and Zhang, L and Li, J and Du, G and Lv, X and Chen, J and Ledesma-Amaro, R and Liu, L},
doi = {nar/gkae228},
journal = {Nucleic Acids Research},
pages = {4739--4755},
title = {Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor},
url = {},
volume = {52},
year = {2024}

RIS format (EndNote, RefMan)

AB - Mutagenesis driving genetic diversity is vital for understanding and engineering biological systems. However, the lack of effective methods to generate in-situ mutagenesis in multiple genomic loci combinatorially limits the study of complex biological functions. Here, we design and construct MultiduBE, a dCas12a-based multiplexed dual-function base editor, in an all-in-one plasmid for performing combinatorial in-situ mutagenesis. Two synthetic effectors, duBE-1a and duBE-2b, are created by amalgamating the functionalities of cytosine deaminase (from hAPOBEC3A or hAIDΔ ), adenine deaminase (from TadA9), and crRNA array processing (from dCas12a). Furthermore, introducing the synthetic separator Sp4 minimizes interference in the crRNA array, thereby facilitating multiplexed in-situ mutagenesis in both Escherichia coli and Bacillus subtilis. Guided by the corresponding crRNA arrays, MultiduBE is successfully employed for cell physiology reprogramming and metabolic regulation. A novel mutation conferring streptomycin resistance has been identified in B. subtilis and incorporated into the mutant strains with multiple antibiotic resistance. Moreover, surfactin and riboflavin titers of the combinatorially mutant strains improved by 42% and 15-fold, respectively, compared with the control strains with single gene mutation. Overall, MultiduBE provides a convenient and efficient way to perform multiplexed in-situ mutagenesis.
AU - Wu,Y
AU - Li,Y
AU - Liu,Y
AU - Xiu,X
AU - Liu,J
AU - Zhang,L
AU - Li,J
AU - Du,G
AU - Lv,X
AU - Chen,J
AU - Ledesma-Amaro,R
AU - Liu,L
DO - nar/gkae228
EP - 4755
PY - 2024///
SN - 0305-1048
SP - 4739
TI - Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor
T2 - Nucleic Acids Research
UR -
UR -
UR -
UR -
VL - 52
ER -