In Vitro Self-Circularization Methods Based on Self-Splicing Ribozyme.

Int J Mol Sci

R&D Center, Rznomics Inc., Seongnam 13486, Republic of Korea.

Published: August 2024


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

In vitro circular RNA (circRNA) preparation methods have been gaining a lot of attention recently as several reports suggest that circRNAs are more stable, with better performances in cells and in vivo, than linear RNAs in various biomedical applications. Self-splicing ribozymes are considered a major in vitro circRNA generation method for biomedical applications due to their simplicity and efficiency in the circularization of the gene of interest. This review summarizes, updates, and discusses the recently developed self-circularization methods based on the self-splicing ribozyme, such as group I and II intron ribozymes, and the pros and cons of each method in preparing circRNA in vitro.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11394858PMC
http://dx.doi.org/10.3390/ijms25179437DOI Listing

Publication Analysis

Top Keywords

self-circularization methods
8
methods based
8
based self-splicing
8
self-splicing ribozyme
8
biomedical applications
8
vitro
4
vitro self-circularization
4
ribozyme vitro
4
vitro circular
4
circular rna
4

Similar Publications

A Computationally Optimized Ribonucleic Acid Circularization Strategy without Byproducts.

J Am Chem Soc

August 2025

Engineering Research Center of Advanced Rare Earth Materials (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing 100084, China.

Circular mRNA (mRNA) exhibits promising potential in mRNA therapy due to its increased stability and extended duration of protein translation, which has sparked an urgent demand for efficient methods to prepare circular RNAs . Here, we present a versatile self-circularization strategy that employs simple motifs to synthesize circular RNAs, achieving robust efficiencies for sequences ranging from dozens to thousands of nucleotides. By leveraging an automated computational program, we optimized highly specific lock-key structures to maximize circularization efficiency, particularly for long RNA substrates.

View Article and Find Full Text PDF

In vitro circular RNA (circRNA) preparation methods have been gaining a lot of attention recently as several reports suggest that circRNAs are more stable, with better performances in cells and in vivo, than linear RNAs in various biomedical applications. Self-splicing ribozymes are considered a major in vitro circRNA generation method for biomedical applications due to their simplicity and efficiency in the circularization of the gene of interest. This review summarizes, updates, and discusses the recently developed self-circularization methods based on the self-splicing ribozyme, such as group I and II intron ribozymes, and the pros and cons of each method in preparing circRNA in vitro.

View Article and Find Full Text PDF

Circular RNA (circRNA) has various advantages over linear mRNA that is gaining success as a new vaccine and therapeutic agent. Thus, circRNA and its engineering methods have attracted attention recently. In this study, we developed a new circRNA engineering method by end-to-end self-targeting and splicing (STS) reaction using group I intron ribozyme.

View Article and Find Full Text PDF

Circularization of for Genotypic Bedaquiline Resistance Testing of Mycobacterium tuberculosis.

Microbiol Spectr

March 2023

Division of Pulmonary and Critical Care Medicine, Zuckerberg San Francisco General Hospital and Trauma Centre, University of California, San Francisco, California, USA.

Circular DNA offers benefits over linear DNA in diagnostic and field assays, but currently, circular DNA generation is lengthy, inefficient, highly dependent on the length and sequence of DNA, and can result in unwanted chimeras. We present streamlined methods for generating PCR-targeted circular DNA from a 700 bp amplicon of , the high GC content (65%) gene implicated in Mycobacterium tuberculosis bedaquiline resistance, and demonstrate that these methods work as desired. We employ self-circularization with and without splints, a Gibson cloning-based approach, and novel 2 novel methods for generating pseudocircular DNA.

View Article and Find Full Text PDF

Easy Hi-C: A Low-Input Method for Capturing Genome Organization.

Methods Mol Biol

November 2022

Department of Genetics and Genome Sciences, School of Medicine, Case Western Reserve University, Cleveland, OH, USA.

Chromosome conformation capture technology and its derivatives have been widely used to study genome organization. Among them, Hi-C (chromosome conformation capture coupling with high-throughput sequencing) is popular in dissecting chromatin architecture on the genome-wide level. However, the intrinsic limitations prevent its application when it comes to rare samples.

View Article and Find Full Text PDF