This disclosure relates to the genetic modification of DNMT3A gene in immune cells. In certain embodiments, the modified immune cells may be used in adoptive T cells therapies to enhance immune responses against cancer or chronic infections. In certain embodiments, the disclosure relates to deleting, changing, or inserting nucleotides within the DNMT3A gene in immune cells, e.g., human CD8 T cells, such that the DNMT3A gene product does not function for methylation. In certain embodiments, modification of the DNMT3A gene provides an improvement in antigen-specific T cells functions and/or an enhancement of the longevity of the cells.

The disclosure provides constructs comprising a first fusion protein, a second fusion protein, and a linker, wherein the first fusion protein and the second fusion protein each include an affinity reagent and a reactive enzyme, and the linker includes a first and second functional groups specific for irreversibly inhibiting the first and second fusion protein reactive enzymes. The disclosure further provides a method including (a) contacting a first fusion protein including an affinity reagent and a reactive enzyme with a linker including a functional group specific for irreversibly inhibiting the first fusion protein reactive enzyme thereby coupling the first fusion protein and the linker, and (b) contacting a second fusion protein including an affinity reagent and a reactive enzyme with the linker, the linker including a functional group specific for irreversibly inhibiting the second fusion protein reactive enzyme thereby coupling the second fusion protein and the linker.

Methods for reactivating genes on the inactive X chromosome that include administering one or both of a DNA methyltransferase (DNMT) Inhibitor and/or a topoisomerase inhibitor, e.g., etoposide and/or 5-azacytidine (aza), optionally in combination with an inhibitor of XIST RNA and/or an Xist-interacting protein, e.g., a chromatin-modifying protein, e.g., a small molecule or an inhibitory nucleic acid (such as a small inhibitory RNA (siRNAs) or antisense oligonucleotide (ASO)) that targets XIST RNA and/or a gene encoding an Xist-interacting protein, e.g., a chromatin-modifying protein.

The present disclosure provides compositions of CRISPR associated (Cas) effector proteins fused to partner proteins. Compositions typically comprise a guide nucleic acid. Also disclosed are the methods and systems for detecting and modifying target nucleic acids using the same. The cells, progenies thereof, and populations thereof produced by the compositions, methods, or systems provided herein are also described.

The disclosure relates to methods and compositions for reactivating a silenced FMR1 gene. In some aspects, methods described by the disclosure are useful for treating a FMR1-inactivation-associated disorder (e.g., fragile X syndrome).

The present disclosure relates to a method for increasing lipid content in microorganisms. The method comprises decreasing the expression of molecules involved in the protein synthesis to decrease protein synthesis and thereby increase lipid synthesis in the microorganisms. The present disclosure also provides a modified microorganism having increased lipid content.

Methods and systems for obtaining inhibitors of human DNA methyltransferase 1 (DNMT1) are disclosed where the methods involve designing compounds that resemble the DNMT1 transition state.

The present disclosure provides a method of enzymatically methylating a DNA by combining a source DNA encoding a pro-apoptotic protein, a determinant protein, or a functional fragment thereof, an extracellular methylation enzyme and an enzymatic substrate in an amount sufficient to allow methylation of at least one CpG site on the source DNA and then incubating the reaction sample at a temperature and for a time sufficient to obtain an enzymatically methylated DNA having a specific methylation level or a specific methylation pattern. The disclosure further provides enzymatically methylated DNA, pharmaceutical compositions containing enzymatically methylated DNA, and methods of treatment using such enzymatically methylated DNA or pharmaceutical compositions.

The present disclosure provides compositions with a modulating gene expression and methods for modulating transcription.

RNA molecules inhibiting a DNMT and methods and compositions incorporating or generating the RNA molecules are described.