The design of this experiment requires comprehensive consideration of genetic biology, neuroscience, ethics, and technical feasibility. The following is a step-by-step comprehensive experimental design plan, focusing on scientific rigor and ethical safety： --- ###**Experimental goal** Through targeted genetic modification (such as CRISPR-Cas9) combined with optogenetic technology, human vision (such as rod/cone sensitivity), hearing (cochlear hair cell function) and neural response speed (synaptic transmission efficiency) are enhanced. --- ###**Stage 1: In vitro cell experiment** 1. **Target gene selection**： -**Vision**: Modify the rhodopsin gene (*RHO*) or the gene that regulates the photoreceptor pigment of cones (such as *OPN1SW*) to enhance sensitivity to low light or specific wavelengths. -**Hearing**: Target the mechanosensitive channel genes (such as *TMC1*) of the hair cells of the cochlea to improve the efficiency of sound wave conversion. -**Nerve conduction**: Overexpression of voltage-gated sodium channel genes (such as *SCN1A*) in neurons, or regulation of the density of AMPA receptors (*GRIA1*) in the postsynaptic membrane. 2. **CRISPR editing verification**： -Introduce editing in human retinal pigment epithelial cells (RPE) and spiral ganglion cells (SGN) to verify gene expression efficiency and functional changes (such as calcium imaging to detect signal strength/speed). 3. **Safety screening**： -Whole genome sequencing detects off-target effects and evaluates cell survival and apoptosis markers (such as Caspase-3). --- ###**Stage 2: Organoids and animal models** 1. ** Organoid test**： -Use human retinal organoids and cochlear organoids to test the response to light/sound stimuli after gene editing (such as multi-electrode arrays to record electrical signals). 2. **Transgenic mouse experiment**： -Generate mice carrying the target gene and verify the phenotype through the following tests： -**Behavior**: Visual water maze task, auditory fright reflex incubation period. -**Physiology**: The incubation period and amplitude of visual evoked potential (VEP) and auditory brainstem response (ABR). -**Side effects**: Monitor the threshold of epilepsy (risk of neurotransmission abnormalities). --- ###**Stage 3: Pre-clinical trial of nonhuman primates (NHP)** 1. **Partial delivery technology**： -Directional delivery of editing components to the retina and cochlea through AAV vectors to avoid systemic exposure. - Use nuclear magnetic resonance (fMRI) to detect changes in the response of the central nervous system. 2. **Cognitive and ethical assessment**： -Test NHP's performance in complex tasks (such as dynamic visual tracking, voice positioning), while monitoring anxiety/aggressive behavior. --- ###**Stage 4: Controlled human clinical Trial** 1. **Subject selection**： -Recruit adult volunteers (18-40 years old), divided into healthy group and sensory degeneration group, double-blind randomized controlled design. 2. **Delivery and monitoring**： -Subretinal injection or round window membrane injection of AAV carrier. -Regular evaluation： -**Subjective**: visual contrast sensitivity (Freiburg test), pure tone hearing threshold. -**Objective**: ERG (retinal electrography) and OAE (otoacoustic emission) detect sensory cell function. -**Neural response**: EEG measures the incubation period of P300 waves (cognitive processing speed). 3. **Long-term follow-up**： -Follow up for more than 5 years to screen for immune response, gene editing stability and potential delayed side effects. --- ###**Ethics and Risk Control** 1. **Approved by the Ethics Committee**： -Comply with the "Declaration of Helsinki", ensure the informed consent of subjects, and clarify the exit mechanism. 2. **Risk mitigation**： -Use suicide genes (such as HSV-TK) as a safety switch to remove edited cells if necessary. -Strictly limit the scope of editing (avoid germ cell or whole brain editing). --- ###**Expected results and challenges** -**Success indicators**: The sensory threshold is reduced by 20%, and the neural response speed is increased by 15% (vs. the control group). -**Potential problems**: Individual differences in gene editing efficiency, immune rejection, sensory overload (such as light/sound sensitivity). -**Alternative **: If the risk of gene editing is too high, you can turn to non-invasive neural regulation (such as transcranial magnetic stimulation). --- ###**Technical roadmap** ```mermaid graph LR A [Target gene screening]--> B [in vitro cell editing] B-->C[organoid verification] C-->D[mouse model] D--> E[NHP security] E-->F[human Phase I clinical] ``` The experiment requires interdisciplinary cooperation (molecular biology, neuroengineering, bioethics), and safety and effectiveness are verified in stages. The ultimate goal is to provide treatment options for patients with sensory impairments, not for healthy people.