Rosier, J. | Search bioRxiv

bioRxiv2025

ZIP editors for efficient and versatile CRISPR-Cas9 precise genome editing

Precise genome editing technologies create the potential for genetic studies and innovative gene therapies. Here we present new CRISPR-Cas9 tools, named ZIP CRISPR, loaded with a single-stranded oligodeoxynucleotide (ssODN) template on the Cas ribonucleoprotein complex. The ssODN template is annealed to an extended guide RNA (gRNA) allowing its nuclear delivery at the right place, i.e. the targeted DNA cut, and at the right time. This new template import system is easy-to-design, easy-to-use, inexpensive and versatile. It increases homology-directed repair (HDR) editing efficiency using Cas9 nuclease up to 12-fold with a mean increase of 5-fold as demonstrated at many loci in many cell types. Based on a heteroduplex gRNA-ssODN, it can also be used with the Cas9 nickase, resulting in HDR editing with minimal InDels, and preventing double-strand break (DSB)-mediated genotoxicity. ZIP CRISPR is a non-viral platform adaptable to targeted DSB (higher HDR editing efficiency) or nick (higher safety) to precisely model and correct a wide range of edits. It is suitable for many biological applications and could be considered for HDR-based gene therapies.

bioengineering↗

bioRxiv2022

The effects of conscious movement processing on the neuromuscular control of posture

Maintaining balance is thought to primarily occur sub-consciously. Occasionally, however, individuals will direct conscious attention towards balance, e.g., in response to a threat to balance. Such conscious movement processing (CMP) increases the reliance on attentional resources and may disrupt balance performance. However, the underlying changes in neuromuscular control remain poorly understood. We investigated the effects of CMP (manipulated using verbal instructions) on neural control of posture in twenty-five adults (11 females, mean age = 23.9, range = 18-33). Participants performed 90-second, bipedal stance balance trials in high- and low-CMP conditions, during both stable (solid surface) and unstable (foam) task conditions. Postural sway amplitude, frequency and complexity were used to assess postural control. Surface EMG was recorded bilaterally from lower leg muscles (Soleus, Tibialis Anterior, Gastrocnemius Medialis, Peroneus Longus) and intermuscular coherence (IMC) was assessed for 12 muscle pairs across four frequency bands. We observed significantly increased sway amplitude, and decreased sway frequency and complexity in the high- compared to the low-CMP conditions. All sway variables increased in the unstable compared to the stable conditions. We observed reduced beta band IMC between several muscle pairs during high- compared to low-CMP, but these findings did not remain significant after controlling for multiple comparisons. Finally, IMC significantly increased in the unstable conditions for most muscle combinations and frequency bands. In all, results tentatively suggest that CMP-induced changes in sway outcomes may be facilitated by reduced beta-band IMC, but these findings need to be replicated before they can be interpreted more conclusively.

neuroscience↗