Related Experiment Video
Updated: Feb 9, 2026
07:17
Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
Published on: December 16, 2022
4.1K
DMSO increases efficiency of genome editing at two non-coding loci.
George Stratigopoulos1,2, Maria Caterina De Rosa1,2,3, Charles A LeDuc1,2,4
1Department of Pediatrics, Columbia University, New York, NY, United States of America.
Plos One
|June 5, 2018
Summary
Dimethyl sulfoxide (DMSO) treatment significantly enhances CRISPR/Cas9 gene editing efficiency for non-coding DNA. This simple method increases single base-pair substitution rates up to 10-fold in human embryonic stem cells without off-target effects.
Area of Science:
- Molecular Biology
- Genetics
- Stem Cell Research
Background:
- CRISPR/Cas9 is a powerful genome editing tool, but its efficiency in non-coding DNA is limited.
- Precise editing of non-coding regions, like intronic single nucleotide polymorphisms (SNPs), is challenging.
Purpose of the Study:
- To improve the efficiency of CRISPR/Cas9-mediated single base-pair substitutions in non-coding DNA.
- To investigate the effect of dimethyl sulfoxide (DMSO) on CRISPR/Cas9 editing efficiency in human embryonic stem cells (ESCs).
Main Methods:
- Human ESCs were treated with 1% DMSO prior to CRISPR/Cas9 delivery.
- Targeted single base-pair substitutions were introduced at intronic SNPs within the FTO locus.
- Off-target effects and chromosomal stability were assessed post-treatment.
Main Results:
- DMSO treatment increased the efficiency of editing non-coding SNPs by up to 10-fold.
- No significant off-target mutations or compromise in chromosomal stability were observed.
- The method allowed for precise editing without altering adjacent non-coding sequences.
Conclusions:
- Pre-treatment with DMSO is a simple and effective strategy to enhance CRISPR/Cas9 editing efficiency for non-coding DNA.
- This approach offers a promising method for precise genetic modifications in ESCs without introducing unwanted mutations.
Related Concept Videos
01:28CRISPR/Cas9 Genome Editing
1.9K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.9K
02:23RNA Editing
9.9K
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...
9.9K
02:39lncRNA - Long Non-coding RNAs
10.0K
In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
10.0K
02:39lncRNA - Long Non-coding RNAs
3.7K
3.7K
03:21Genome Size and the Evolution of New Genes
9.2K
While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
9.2K
02:02Genomics
40.8K
Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
40.8K