The present disclosure provides for systems, methods, and compositions for targeting nucleic acids. In particular, the invention provides mutated Cas13 proteins and their use in modifying target sequences as well as mutated Cas13 nucleic acid sequences and vectors encoding mutated Cas13 proteins and vector systems or CRISPR-Cas13 systems.

Reagents and methods to cloak and uncloak RNA polymers and applications thereof are provided. Photocloaking molecules are used to label RNA polymers. Radiant energy is used to remove photoreleaseable protecting adducts and revert a RNA polymer to its native form.

Provided herein are compositions and methods for increasing editing efficiency of a target nucleic acid. A composition may comprise a guide nucleic acid, a Cas9 nickase, or a reverse transcriptase. The reverse transcriptase may be fused to the Cas9 nickase. The reverse transcriptase may heterodimerize with the Cas9 nickase. The reverse transcriptase may bind to a guide nucleic acid. The reverse transcriptase may be engineered to increase processivity. The guide nucleic acid may be engineered to facilitate synthesis or editing of a sequence. The guide nucleic acid, Cas9 nickase, and reverse transcriptase may be engineered to fit within AAV vectors. The guide nucleic acid may comprise a region that binds to another region on the guide nucleic acid to improve gene editing.

There is a method for selective translation of a desired protein. The method has the steps of (a) providing a modified nucleic acid enzyme, including two half cores of a minimized 9DB1 deoxyribozyme split between nucleotides 35 and 39, wherein each half core includes a pendant assembly arm of a strand of nucleic acids extending therefrom and a separate, pendant binding arm extending therefrom of a strand of nucleic acids; (b) binding a nucleic acid ligand to each of the two assembly arms to form an intermediate; (c) binding the intermediate to (i) a first substrate of ribonucleic acid sequences capped at one end, (ii) a second substrate of a strand of ribonucleic acids having a 5′ triphosphate region at one end and a region of polyadenylated nucleotides at the other end and wherein the second substrate codes for the desired protein, (iii) join the two half cores to form a core in order to form a ligated product; and (d) allowing the translation for the desired protein to proceed from the ligated product. There is another method for selective translation of a desired protein. There is also a modified nucleic acid enzyme.

The present invention relates to signalling catalytic nucleic acid nanostructures that are responsive to polymerase activity, methods of their use, devices and kits comprising the same. More specifically, the present invention provides a catalytic signalling nanostructure comprising a DNAzyme/RNAzyme, such as G-quadruplex hemin, and a polymerase—responsive element. Polymerase elongation of the polymerase-responsive element eliminates catalytic activity of the DNAzyme/RNAzyme. The catalytic nucleic acid nanostructure can be used alone or paired with a target recognition nanostructure which can transduce molecular signals into polymerase activity, in an integrated circuit.

The present disclosure relates to methods for transiently activating temperature-sensitive agents in one or more cells, for example by contacting one or more cells with a temperature-sensitive agent and transiently incubating the cells at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Additionally, the present disclosure relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent and then lowering the subject's core body temperature to a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. The disclosure also relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent, maintaining the subject's surface body temperature at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Further disclosed are methods of treating a subject with a temperature-sensitive therapeutic agent.

The invention relates to the field of molecular biology and cell biology. More specifically, the invention relates to CRISPR guide-RNA expression strategies for multiplex genome engineering.

The present disclosure provides compositions of matter, methods and instruments for editing nucleic acids in live yeast cells.

A ribozyme comprising a target-binding domain, a polynucleotide encoding the same, a kit comprising the same, and uses thereof.

The present disclosure relates to methods for transiently activating temperature-sensitive agents in one or more cells, for example by contacting one or more cells with a temperature-sensitive agent and transiently incubating the cells at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Additionally, the present disclosure relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent and then lowering the subject's core body temperature to a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. The disclosure also relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent, maintaining the subject's surface body temperature at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Further disclosed are methods of treating a subject with a temperature-sensitive therapeutic agent.