Howard Hughes Medical Institute

E.i. Dupont De Nemours and Company

Pioneer Hi-Bred International, Inc.

The Brigham and Women's Hospital, Inc.

Duke University

University of California

New York University

Loren Lee Looger

Eric R Schreiter

Jonathan S Marvin

Douglas S Kim

Karel Svoboda

Michael W Lassner

Kevin E McBride

Brian McGonigle

Richard Theodore Lee

Rex Kerr

Benjamin F Fosque

David V Schaffer

Homme W Hellinga

Bum-Yeol Hwang

Joshua Dudman

Alla Karpova

Dougal Gowanlock Robinson Tervo

Adam Hantman

Kimberly Ritola

Sarada Viswanathan

Scott Sternson

Peter Lee

Dmitry Rinberg

Lin Tian

Yan Zhang

Jeremy P Hasseman

Ilya Kolb

Douglas Kim

The present disclosure provides, inter alia, genetically encoded recombinant peptide biosensors comprising analyte-binding framework portions and signaling portions, wherein the signaling portions are present within the framework portions at sites or amino acid positions that undergo a conformational change upon interaction of the framework portion with an analyte.

Genetically encoded biosensors

Genetically encoded calcium indicator (GECI) polypeptides and the nucleic acid molecules encoding such polypeptides are provided.

Genetically encoded calcium indicators (GECIs) and methods of making and using

The present disclosure provides AAV variants that exhibit a preference for retrograde movement in neurons and methods of using such variants.

Variant adeno-associated viruses and methods of using

Genetically encoded calcium indicator (GECI) polypeptides and the nucleic acid molecules encoding such polypeptides are provided. In addition, methods of using such nucleic acids and polypeptides in methods of screening for agonists or antagonists of G-protein coupled receptor (GPCR) or ion channels and methods of monitoring neural activity also are provided.

Genetically encoded calcium indicators and methods of use

Biosensors are made by attaching covalently or non-covalently at least one reporter group to one or more specific positions of a bacterial periplasmic binding protein (bPBP). Upon binding of ligand to the biosensor, there is a change in the signal transduced by the reporter group.

Biosensor

The presently-disclosed subject matter includes fluorescent protein-based indicators for detecting ions, small molecule analytes, or combinations thereof. In some embodiments the indicators include a polypeptide, which itself includes a fluorescent polypeptide, a compound-binding polypeptide, and a polypeptide target of the compound-binding polypeptide. In some embodiments the polypeptide includes an EosFP polypeptide, a calmodulin polypeptide, and a M13 polypeptide, or fragments and/or variants thereof. The presently-disclosed subject matter also includes methods for detecting calcium in a sample with embodiments of the present polypeptides. In some embodiments the present indicators experience a permanent shift from green to red fluorescent when exposed to an detecting substance, such as calcium.

Methods of using fluorescent protein-based indicators

Protein indicators useful for calcium imaging, in particular, red genetically-encoded calcium indicators (GECIs) disclosed herein rival best-of-class green GECIs in terms of sensitivity for detecting neural activity, and can be monitored in vivo. The presently-disclosed subject matter further includes a method of monitoring cell activity comprising stimulating a cell comprising a red GECI polypeptide; and detecting fluorescence emitted by the cell.

Red genetically encoded calcium indicators and methods of use

This disclosure provides methods for identification of olfactory sensosry neurons (OSN) that recognize specific odorants. The method comprises introducing into the OSN, a permanent activity marker which exhibits a detectable and permanent change upon activation of the neuron in response to an odorant and exposure to an exteranal stimulus, such as light. The OSN can be isolated and its receptor characterized.

Identification of olfactory receptors sensitive to different odorants

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Madison, AL, United States of America

Loren Lee Looger