Structure-guided SCHEMA recombination generates diverse chimeric channelrhodopsins
Abstract
Critical for regulating cell function, integral membrane proteins (MPs) are key engineering targets. MP engineering is limited because these proteins are difficult to express with proper plasma membrane localization in heterologous systems. We investigate the expression, localization, and light-induced behavior of the light-gated MP channel, channelrhodopsin (ChR), because of its utility in studying neuronal circuitry. We used structure-guided SCHEMA recombination to generate libraries of chimeric ChRs that are diverse in sequence yet still capable of efficient expression, localization, and useful light-induced functionality. The conservative nature of recombination generates unique protein sequences that tend to fold and function. Recombination is also innovative: chimeric ChRs can outperform their parents or even exhibit properties not known in natural ChRs.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 10, 2017
- Source ID
- 10.1073/pnas.1700269114
Entities
People
- Austin J. Rice
- Claire N Bedbrook
- Emily M. Leproust
- Frances Arnold
- Kevin K. Yang
- Siyuan Chen
- Viviana Gradinaru
- Xiaozhe Ding
Organizations
- Army Research Office
- California Institute of Technology
- National Center for Research Resources
- National Institute of General Medical Sciences
- National Institute of Mental Health
- Twist Bioscience