Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

5.9K
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...
5.9K
Experimental RNAi02:15

Experimental RNAi

6.0K
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
6.0K
Alternative RNA Splicing02:18

Alternative RNA Splicing

20.9K
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
20.9K
Riboswitches01:56

Riboswitches

8.0K
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
8.0K
siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

16.5K
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
16.5K
RNA Interference01:23

RNA Interference

25.9K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
25.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Multidimensional Motion and Manipulation of Optothermal Bubble Microrobots in Dimethyl Silicone Oil via Remote Laser Control.

ACS applied materials & interfaces·2026
Same author

Natto May Alleviate Retinoic Acid-Induced Osteoporosis by Activating Gut Microbiota-Bile Acid Axis and OPG/RANKL Signaling Pathway.

Nutrients·2026
Same author

PYCR1 modulates ferroptosis and malignant phenotypes of clear cell renal cell carcinoma through proline biosynthesis.

Molecular biology reports·2026
Same author

Safety Profile of COVID-19 Vaccines in HIV Patients Undergoing ART and Their Impact on Immune Recovery and HIV Reservoirs.

Infectious diseases & immunity·2026
Same author

Association between CHRNA5 polymorphisms and Parkinson's disease in Northern Chinese Han population: a case-control study.

Annals of human biology·2026
Same author

The selenium-enriched <i>Rhodotorula mucilaginosa</i> JAASRY1 improved oxidative stress during the aging process via the gut-liver-brain axis.

Frontiers in microbiology·2026
Same journal

Correction to 'New origin firing is inhibited by APC/CCdh1 activation in S-phase after severe replication stress'.

Nucleic acids research·2026
Same journal

VeloRM: disentangling pre- and post-splicing RNA modification dynamics at single-cell resolution.

Nucleic acids research·2026
Same journal

Accessibility of telomeric overhangs to stabilizing small-molecule ligands.

Nucleic acids research·2026
Same journal

Multivalent interactions mediate SNAIL transcription factor stimulation of the nucleosome deacetylase activity of the CoREST complex.

Nucleic acids research·2026
Same journal

Genome-wide mapping of DNA G-quadruplexes in Trypanosoma brucei chromatin reveals enrichment in coding regions and transcription start sites.

Nucleic acids research·2026
Same journal

Correction to 'The Gene Ontology knowledgebase in 2026'.

Nucleic acids research·2026
See all related articles

Related Experiment Video

Updated: Jun 2, 2025

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
09:26

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

4.1K

A trigger-inducible split-Csy4 architecture for programmable RNA modulation.

Lihang Zhang1,2,3,4, Xinyuan Qiu5,6, Yuting Zhou1,3,4

  • 1School of Medicine, Westlake University, No. 18 Shilongshan Road, Xihu District, Hangzhou, Zhejiang, 310024, China.

Nucleic Acids Research
|January 16, 2025
PubMed
Summary
This summary is machine-generated.

We developed a split-CRISPR-Cas system (split-Csy4) to minimize unintended RNA cleavage in gene therapy. This engineered system shows reduced off-target effects compared to full-length Csy4, enabling safer gene regulation.

More Related Videos

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

1.7K
An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling
08:34

An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling

Published on: December 18, 2017

6.6K

Related Experiment Videos

Last Updated: Jun 2, 2025

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
09:26

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

4.1K
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

1.7K
An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling
08:34

An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling

Published on: December 18, 2017

6.6K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Therapy

Background:

  • CRISPR-derived endoribonuclease Csy4 is utilized for transgene expression control.
  • Potential off-target RNA cleavage by Csy4 presents a safety concern for therapeutic applications.
  • The exploration of Csy4's adverse effects and development of safer alternatives is crucial.

Purpose of the Study:

  • To engineer and validate a split-Csy4 system for enhanced safety and efficacy in vivo.
  • To assess the impact of split-Csy4 on the endogenous transcriptome compared to full-length Csy4.
  • To develop inducible gene switches using the split-Csy4 module for precise gene regulation.

Main Methods:

  • Deconstructing the Csy4 enzyme into two independent protein moieties.
  • Reconstituting catalytic activity through various protein dimerization systems.
  • Evaluating transcriptome perturbation in human cells upon introduction of split-Csy4 versus full-length Csy4.
  • Engineering inducible gene switches regulated by grazoprevir.

Main Results:

  • The split-Csy4 architecture significantly reduced perturbation of the endogenous transcriptome in human cells.
  • Full catalytic activity of Csy4 was restored using inducible dimerization systems.
  • Inducible CRISPR- and translation-level gene switches were successfully engineered using the split-Csy4 module.

Conclusions:

  • The split-Csy4 system offers a safer alternative to full-length Csy4 for controlling transgene expression.
  • This engineered system minimizes off-target RNA cleavage, addressing a key safety concern.
  • The developed inducible gene switches provide a valuable resource for Csy4-based biomedical research and clinical applications.