Fanzor is a eukaryotic programmable RNA-guided endonuclease
RNA-guided systems, which use complementarity between a guide RNA and target nucleic acid sequences for recognition of genetic elements, have a central role in biological processes in both prokaryotes and eukaryotes.
Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells
We describe a strategy to boost the efficiency of gene editing via homology-directed repair (HDR) by covalently modifying the template DNA with interstrand crosslinks.
Targeting TBK1 to overcome resistance to cancer immunotherapy
Here we identify the innate immune kinase TANK-binding kinase 1 (TBK1) as a candidate immune-evasion gene in a pooled genetic screen. Using a suite of genetic and pharmacological tools across multiple experimental model systems, we confirm a role for TBK1 as an immune-evasion gene.
Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases
We present programmable addition via site-specific targeting elements (PASTE), which uses a CRISPR–Cas9 nickase fused to both a reverse transcriptase and serine integrase for targeted genomic recruitment and integration of desired payloads.
Real-Time Culture-Independent Microbial Profiling Onboard the International Space Station Using Nanopore Sequencing
Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS.
Detection of SARS-CoV-2 with SHERLOCK One-Pot Testing
Here, we describe a simple test for detection of SARS-CoV-2. The sensitivity of this test is similar to that of reverse-transcription–quantitative polymerase-chain-reaction (RT-qPCR) assays. STOP (SHERLOCK testing in one pot) is a streamlined assay that combines simplified extraction of viral RNA with isothermal amplification and CRISPR-mediated detection.
A 5-min RNA preparation method for COVID-19 detection with RT-qPCR
RNA extraction has become a bottleneck for detection of COVID-19, in part because of reagent shortages. We present here a rapid protocol that circumvents the need for RNA extraction that is compatible with RT-qPCR-based detection methods.
Genome engineering using the CRISPR-Cas9 system
Here we describe a set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies.