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Genetic Engineering Publications - GEG Tech top picks
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“CRISPR” validation of recessive brain cancer genes in vivo
BigField GEG Tech's insight:
In this editorial, the authors describe the power of the CRISPR system to enhance the screening of oncogenes and the understanding of cancer in the brain. 
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Oncogenic driver mutations are identified in single cells by a transposon-based sequencing method.
BigField GEG Tech's insight:
Transposon-based mutagenesis allows the identification of early cancer drivers, but current sequencing methods have limitations that prevent single-cell analysis. Here, the scientists report a liquid-phase, capture-based sequencing and bioinformatics pipeline, Sleeping Beauty(SB) capture hybridization sequencing (SBCapSeq), that facilitates sequencing of transposon insertion sites from single tumor cells in a SB mouse model of myeloid leukemia (ML). SBCapSeq analysis of just 26 cells from one tumor revealed the tumor's major clonal subpopulations, enabled detection of clonal insertion events not detected by other sequencing methods and led to the identification of dominant subclones, each containing a unique pair of interacting gene drivers along with three to six cooperating cancer genes with SB-driven expression changes.
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BigField GEG Tech's insight:
The authors describe an efficient method to induce specific chromosomal rearrangements in vivo using viral-mediated delivery of the CRISPR/Cas9 system to somatic cells of adult animals. They apply it to generate a mouse model of Eml4–Alk-driven lung cancer.
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Here, the scientists examined the piggyBac (PB) transposon as an alternative vehicle to deliver a guide RNA (gRNA) library for in vivo screening. Through hydrodynamic tail vein injections, they delivered a PB-CRISPR library into mouse liver. Rapid tumor formation could be observed in less than 2 mo. By sequencing analysis of PB-mediated gRNA insertions, they identified corresponding genes mediating tumorigenesis. Their results demonstrate that PB is a simple and nonviral choice for efficient in vivo delivery of CRISPR libraries for phenotype-driven screens.