PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells.

Prime editing is a recently developed CRISPR/Cas9 based gene engineering tool that allows the introduction of short insertions, deletions, and substitutions into the genome. However, the efficiency of prime editing, which typically achieves editing rates of around 10%-30%, has not matched its versatility. Here, we introduce the prime editor activity reporter (PEAR), a sensitive fluorescent tool for identifying single cells with prime editing activity. PEAR has no background fluorescence and specifically indicates prime editing events. Its design provides apparently unlimited flexibility for sequence variation along the entire length of the spacer sequence, making it uniquely suited for systematic investigation of sequence features that influence prime editing activity. The use of PEAR as an enrichment marker for prime editing can increase the edited population by up to 84%, thus significantly improving the applicability of prime editing for basic research and biotechnological applications.

Gender inequalities in the treatment of osteoporosis.

INTRODUCTION: Our aim was to perform both gender- and age-specific analysis regarding the utilisation of anti-osteoporotic drugs in Hungary, between 2007 and 2011, and to compare our results with other European countries. METHODS: The database of the Hungarian National Health Insurance Fund was screened for anti-osteoporotic medications, covering 100% of the Hungarian population (10 million people). ICD coding system (International Classification of Diseases) and WHO ATC/DDD methodology were used for medication screening and analysis. RESULTS: In Hungary, the total bisphosphonate use was 6.66 DDD/TID (Defined Daily Dose/1000 inhabitants/day) in 2007, and 6.22 DDD/TID in 2011; the rate of bisphosphonate combinations slightly increased from 1.60 to 2.81 DDD/TID. The total vitamin D use almost doubled (13.73 DDD/TID in 2011), while the calcium supplementation tripled (4.47 DDD/TID in 2011), and so did the strontium ranelate utilisation (0.70 DDD/TID in 2011) within the investigated time period. Denosumab consumption was marginal. Male patients were disproportionately, 10-20 times undertreated in all age groups, and treatment choice was restricted among men. Several differences were seen in our results compared to those in Baltic countries, Finland and in Norway. CONCLUSIONS: Men were significantly undertreated in all age groups, compared to women. The 10 to 20-fold difference calls attention to this unrecognised problem.