Biology subjects

Jang, A.-H.

Publications and source records attributed to Jang, A.-H..

3 recordsLinked to original sources

bioRxiv2024

Efficient DNA base editing via an optimized DYW-like deaminase

CRISPR-based cytosine base editors enable precise genome editing without inducing double-stranded DNA breaks, yet traditionally depend on a limited selection of deaminases from the APOBEC/AID or TadA families. Here, we introduce SsCBE, a novel CRISPR-based cytosine base editor utilizing SsdAtox, a DYW-like deaminase derived from the toxin of Pseudomonas syringae. Strategic engineering of SsdAtox has led to remarkable improvements in the base editing efficiency (by up to 8.4-fold) and specificity for SsCBE, while concurrently reducing cytotoxicity. Exhibiting exceptional versatility, SsCBE was delivered and efficiently applied using diverse delivery methods, including the engineered virus-like particles (eVLPs). Its application has enabled targeted cytosine base editing in mouse zygotes and pioneering edits in mitochondrial DNA. The advent of SsCBE marks a significant advancement in the CRISPR toolkit, providing a versatile tool for advanced research and therapeutic strategies.

molecular biology↗

bioRxiv2020

Engineered Prime Editors with PAM flexibility

Although prime editors are a powerful tool for genome editing, which can generate various types of mutations such as nucleotide substitutions, insertions, and deletions in the genome without double-strand breaks or donor DNA, the conventional prime editors are still limited to its target scopes because of the PAM preference of the spCas9 protein. Here, we described the engineered prime editors to expand the range of their target sites using various PAM-flexible Cas9 variants.

bioengineering↗

bioRxiv2018

Cas-CLIP: a method for customizing pooled CRISPR libraries

ABSCTRACTAlthough pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes, library construction for researchers own study is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple, scalable method, called Cas-CLIP, to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. Cas-CLIP is a precise method for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology.

bioengineering↗