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 Experiment Videos

Specific RNA binding by a single C2H2 zinc finger.

W J Friesen1, M K Darby

  • 1Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.

The Journal of Biological Chemistry
|November 1, 2000
PubMed
Summary

Engineered zinc finger proteins specifically bind RNA, including the human immunodeficiency virus Rev responsive element (RRE). These proteins offer insights into RNA-protein interactions and potential therapeutic applications.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

The methylosome, a 20S complex containing JBP1 and pICln, produces dimethylarginine-modified Sm proteins.

Molecular and cellular biology·2001
Same author

SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets.

Molecular cell·2001
Same author

Specific sequences of the Sm and Sm-like (Lsm) proteins mediate their interaction with the spinal muscular atrophy disease gene product (SMN).

The Journal of biological chemistry·2000
Same author

Specific RNA binding proteins constructed from zinc fingers.

Nature structural biology·1998
Same author

The role of zinc finger linkers in p43 and TFIIIA binding to 5S rRNA and DNA.

Nucleic acids research·1998
Same author

Phage display of RNA binding zinc fingers from transcription factor IIIA.

The Journal of biological chemistry·1997

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Zinc finger proteins (ZFPs) can be engineered for specific RNA binding.
  • Previous studies identified ZFPs with high affinity for the human immunodeficiency virus Rev responsive element stem loop IIB (RRE-IIB).

Purpose of the Study:

  • To characterize the RNA binding properties of individual zinc fingers from a previously identified ZFP (RR1).
  • To engineer novel ZFPs with high affinity and specificity for the full human immunodeficiency virus Rev responsive element (RRE234).

Main Methods:

  • Expression and individual assaying of zinc fingers from RR1.
  • In vitro selection and recombination techniques to construct new ZFPs.
  • Determination of dissociation constants (Kd) for RRE-IIB and RRE234 binding.

Main Results:

  • The C-terminal zinc finger of RR1 retained significant RRE-IIB binding affinity, while the N-terminal finger had substantially lower affinity.
  • Engineered ZFPs demonstrated subnanomolar dissociation constants for RRE234.
  • Engineered ZFPs bound RRE-IIB with an affinity two orders of magnitude lower than RRE234, similar to Rev peptide binding.

Conclusions:

  • Single C2H2 zinc fingers can achieve specific RNA binding.
  • ZFP binding to RRE-IIB may mimic Rev peptide interactions.
  • The binding mechanism to the larger RRE234 might differ from RRE-IIB due to structural complexities or altered stem loop IIB conformation within RRE234.

Related Experiment Videos