相关实验视频

Updated: Jul 15, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

用可编程除氨酶对有机细胞DNA进行基编辑

Jin-Soo Kim^1,2, Jia Chen^3,4

¹NUS Synthetic Biology for Clinical & Technological Innovation (SynCTI) and Department of Biochemistry, National University of Singapore, Singapore, Singapore. jskim01@snu.ac.kr.

Nature reviews. Molecular cell biology

|October 4, 2023

概括

新的无CRISPR基编辑器可以精确编辑线粒体和叶绿体中的细胞器DNA. 这些工具为研究遗传疾病,衰老以及开发新疗法和农业应用提供了潜力.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

线粒体和叶绿体拥有自己的基因组,对细胞功能如能源生产和碳固定至关重要.
细胞器DNA的突变与遗传疾病,衰老和癌症等疾病有关.
缺乏针对有机细胞DNA编辑的有效工具,阻碍了研究和治疗开发.

研究的目的:

审查最近在无CRISPR基基编辑器中取得的进展,用于器官DNA编辑.
讨论这些编辑器在线粒体和塑体基因组中的应用.
确定这些编辑技术的局限性,并提出这些编辑技术的未来改进方案.

主要方法:

开发无CRISPR,仅用蛋白质的基编辑器,包括细胞因基编辑器 (DdCBEs) 和腺因基编辑器 (ABEs).
这些编辑器的应用在体外针对性DNA编辑,人类细胞系,动物和植物.
在体外表征可编程除氨酶用于器官DNA操纵.

主要成果:

在各种生物系统中使用新的基根编辑器对有机细胞DNA编辑的演示.
成功应用基基编辑器用于动物中的线粒体DNA编辑和植物中的塑基因组编辑.
识别这些基础编辑工具当前的精度和效率限制.

结论:

没有CRISPR的基编辑器代表了器官DNA编辑的重大进步.
这些工具对基础研究,治疗干预和农业改进充满希望.
需要进一步优化,以提高器官基编辑的精度和效率,以便在更广泛的应用中使用.

更多相关视频

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

相关概念视频

Base Excision Repair

Base Excision Repair

One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...

RNA Editing

RNA Editing

RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...

Overview of DNA Repair

Overview of DNA Repair

In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
Chemically...

DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

Restriction Enzymes

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...