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

Evolving proteins in mammalian cells using somatic hypermutation.

Lei Wang1, Roger Y Tsien

  • 1The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, California 92037, USA. lwang@salk.edu

Nature Protocols
|April 5, 2007
PubMed
Summary
This summary is machine-generated.

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

[Physico-chemical characteristics of ambient particles settling upon leaf surface of six conifers in Beijing].

Ying yong sheng tai xue bao = The journal of applied ecology·2007
Same author

Ribonucleic acid binding protein-mediated regulation of luteinizing hormone receptor expression in granulosa cells: relationship to sterol metabolism.

Molecular endocrinology (Baltimore, Md.)·2007
Same author

Psychological stress-induced oxidative stress as a model of sub-healthy condition and the effect of TCM.

Evidence-based complementary and alternative medicine : eCAM·2007
Same author

Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice.

Planta·2007
Same author

Edge-based scoring and searching method for identifying condition-responsive protein-protein interaction sub-network.

Bioinformatics (Oxford, England)·2007
Same author

[The value of long-term postoperative follow-up after curative resection of lung cancer and common problems associated with it].

Nihon Geka Gakkai zasshi·2007
Same journal

iMUT-seq mapping of DSB-induced mutations with high sensitivity at single-nucleotide resolution.

Nature protocols·2026
Same journal

An assay to quantify sexual commitment and stage conversion in the human malaria parasite Plasmodium falciparum.

Nature protocols·2026
Same journal

Author Correction: Direct inoculation of bioreactor-controlled stirred suspension culture with cryopreserved human pluripotent stem cells.

Nature protocols·2026
Same journal

High-throughput measurements of protein domain functions using magnetic separation.

Nature protocols·2026
Same journal

Inducing physiological polarity and performing gene editing using CRISPR-Cas9 in human trophoblast organoids.

Nature protocols·2026
Same journal

Photocatalytic low-temperature defluorination of PTFE.

Nature protocols·2026
See all related articles

Researchers developed a novel method for protein evolution in mammalian cells using somatic hypermutation (SHM). This technique allows for rapid gene mutation and protein engineering directly within living cells, streamlining the discovery of new protein functions.

Area of Science:

  • Molecular Biology
  • Protein Engineering
  • Cell Biology

Background:

  • Somatic hypermutation (SHM) is a natural process in B-cells that introduces mutations into immunoglobulin genes.
  • Existing in vitro mutagenesis methods are labor-intensive and may not fully capture complex protein interactions.
  • Evolving proteins within living cells offers a more physiologically relevant approach to protein engineering.

Purpose of the Study:

  • To establish a novel method for evolving proteins in living mammalian cells using somatic hypermutation.
  • To demonstrate the feasibility of iterative rounds of mutation, selection, and screening for protein evolution.
  • To bypass traditional in vitro mutagenesis techniques for faster protein engineering.

Main Methods:

  • Utilizing a B-cell line with constitutive immunoglobulin (Ig) V gene hypermutation.

Related Experiment Videos

  • Inducing target gene expression under a controllable promoter to enable SHM-driven mutagenesis.
  • Iterative cycles of gene mutation, protein expression, and cell-based selection or screening for desired phenotypes.

    • Main Results:

    • Successfully evolved a monomeric red fluorescent protein (mRFP1.2) into a far-red emitting mutant (mPlum) in Ramos cells.
    • Demonstrated the accumulation of multiple beneficial mutations through iterative rounds of evolution.
    • Established a rapid workflow with each round of evolution taking approximately 3.5-6 days.

    Conclusions:

    • The described method enables efficient protein evolution directly within living mammalian cells.
    • This approach significantly reduces the time and labor associated with protein engineering.
    • The protocol is adaptable for evolving diverse eukaryotic proteins in various SHM-competent cell types.