Harvard College

Reza Kalhor

George M Church

Henry Hung-yi Lee

Prashant G Mali

Javier Fernandez Juarez

A functional engineered guide RNA sequence is provided including a spacer sequence and a scaffold sequence, wherein the scaffold sequence includes a primer binding site for reverse transcription.

Engineered guide RNA sequences for in situ detection and sequencing

The disclosure provides methods for making a polynucleotide wherein the addition of nucleotides can be physically, chemically and/or enzymatically controlled. The methods include combining a selected nucleotide, cations, an error prone or template independent DNA polymerase at a reaction site including an initiator sequence attached thereto and having a 3' terminal nucleotide, wherein the reaction reagents can be modulated and under conditions that allow covalent addition of one or more of a selected nucleotide to the 3&#x2032; terminal nucleotide such that the selected nucleotide becomes a 3&#x2032; terminal nucleotide, and repeating the addition step until the polynucleotide is formed.

Enzymatic nucleic acid synthesis

A method of processing a collection of nucleic acid sequences is provided including connecting an adaptor to one or more or each nucleic acid sequence in the collection to create a processed nucleic acid template library, wherein the adaptor includes a first DNA sequence encoding a PAM sequence and at least a tracr mate.

Frequency-based modulation of diverse species in a nucleic acid library

The present disclosure provides methods of activating an enzyme, such as error prone or template independent polymerase, using electricity to alter pH of a reaction zone and reaction site from an inactivating pH at which the enzyme is inactive to an activating pH at which the enzyme is active to add a nucleotide to an initiator or growing polymer chain. The activating pH can then be changed back to an inactivating pH and the process repeated as many times as desired to produce a target nucleic acid sequence.

Growing community of inventors

East Boston, MA, United States of America

Reza Kalhor