CRISPR gene editing calls into question results of older genetics studies

[Jason Sheltzer, a cancer biologist at Cold Spring Harbor Laboratory in New York] and his team joined an expanding club of laboratories that have been forced to re-evaluate and repeat experiments, as the spread of CRISPR–Cas9 uncovers potential errors in data collected using older techniques.

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Nathan Lawson, a molecular biologist at the University of Massachusetts Medical School in Worcester, was one of the first to systematically characterize the problem. In 2015, he and his colleagues reported their efforts to compare results from two methods in zebrafish: knocking out genes using a gene-editing technique called zinc finger nucleases, and reducing gene expression using molecular tools called morpholinos. They found that half of the 20 genes they tested yielded different results…

Conflicting results from RNAi and genetic screens do not always mean that one approach was right and the other was wrong, cautions [Michael Bassik, a molecular biologist at Stanford University in California]. Some cells might respond differently to a genetic change that wipes out expression of a gene, as is often the goal with CRISPR–Cas9, compared to how they respond to reducing the expression to very low levels with RNAi.

But often, he adds, the culprit behind the discrepancy can be tracked back to RNAi’s potential for off-target effects. And concerns about that have had researchers flocking to reproduce old results.

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