Roche Molecular Systems, Inc.

Roche Sequencing Solutions, Inc.

Genia Technologies, Inc.

Shawn Lee Suko

Thomas W Myers

Keith Bauer

Fred Lawrence Reichert

Aruna Ayer

Mara Boenitz-Dulat

Ilya Lederman

Eileen Thai

Charles Wayan Schwab

Colin Alexander McGaw

Cleoma Arnold

Barbara Eckert

David Daniel Wunderlich

Bigna Woersdoerfer

David H Gelfand

Ellen H Fiss

Edward Soh Smith

Arkadiusz Bibillo

Joseph San Filippo

Tyler O'Brien Shultz

Rachel Shahinian

Preethi Sarvabhowman

Evan Amato

Nancy J Schoenbrunner

Susanne Stoffel

Olga Budker

Modified or mutant bacterial luciferases having improved activity, as compared to wild type or unmodified bacterial luciferases, are described. The modified or mutant bacterial luciferases display increased light production and/or slower signal decay. Employing these modified or mutant bacterial luciferases improve a luminescence reporter system assay by increasing the detection sensitivity, resulting in improved bioreporter/reporter assays. The invention further provides methods for using the modified or mutant bacterial luciferases, reporter assays using the modified or mutant bacterial luciferases, and kits and articles of manufacture.

Modified/mutant bacterial luciferases

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, I644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

Polymerase variants

The present disclosure provides variant Pol6 polymerase polypeptides, compositions comprising the Pol6 variant polypeptides, and methods for using the variant Pol6 polypeptides for determining the sequencing of nucleic acids, for example, by nanopore sequencing. The variant Pol6 polymerases possess decreased rates of dissociation of template from the polymerase-template complex, which result in increased processivity relative to the parental Pol6 polypeptides from which they are derived.

Pol6 polymerase variants

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, 1644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

Disclosed are DNA polymerases having increased reverse transcriptase efficiency, mismatch tolerance, extension rate and/or tolerance of RT and polymerase inhibitors relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.

DNA polymerases with improved activity

Provided are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of primer extension methods. Also provided are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.

Described herein are variant pol6 polymerase having at least having at least two substitutions, wherein at least one of the substitutions is at position S366 of SEQ ID NO: 2. The disclosed polymerases may be coupled to alpha-hemolysin nanopores, and may be used in nanopore-based sequencing apparatuses.

Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.

Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

Growing community of inventors

Walnut Creek, CA, United States of America

Shawn Lee Suko