• how does CRY actually regulate PER activity?
• without CRY1KO (not CRY2KO), the period is shortened -- so CRY1 must, in some way, lengthen period
• the opposite occurs in CRY2, implying CRY2 shortens period
• CRYDKO leads to complete arrhythmicity
• in CRYDKO, there is still some rhythmicity when the LD cycle is still going, but free-running in LL/DD
• TTFL: PERs and CRYs inhibit their own expression through, in some way, repressing their transcription via BMAL1 and CLK
• KI of venus fluorescence into the same gene for PER2
• fluorescence recovery after photobleaching (FRAP): labelling of all lipids in the PM, blasting part of the PM with a laser to bleach the lipids' fluorescence, seeing how long the lipids take to restore fluorescence intensity
• use FRAP to bleach PER2 to find out how quickly it moves (fast, slow, immobile PER2)
• viruses used as a vector to express transgenes in rescue experiments
• CRY1 and CRY2 ability to rescue differs over time, but both CRYs can rescue expression eventually
• there does not seem to be a significant does dependence in the amount of CRY (even very low levels of CRY are sufficient to translate PER to the nucleus)
• translational switch is a reversing of the genetic code expansion (GCE)
• charging of tRNAs with a "non-canonical amino acid" (ncAA) through engineered tRNA synthetases

• how do orgs correctly regenerate missing tissues?
• it is actually more complicated than these paper let on
• selective cuts could interfere with correct body segment regeneration (this paper justifies the molecular bases of that)
• by KD of certain parts of the Wnt cascade, you can affect the polarity of regeneration response
• non-canonical Wnt signaling: you do not need beta-catenin
• there is most likely a branch point after disheveled, so it can use a signal other than beta-catenin to signal alternative regeneration
• by ridding of beta-catenin, you can induce head growth wherever you want (even at the sides of the animal, not just the ant-/post-)
• under normal development, Wnt does not usually determine ant- v. post- or head v. tail, but it is more complex in its involvement of directional development of diff tissues
• during homeostatic development, beta-catenin also regulates head v. tail actions

/ embryonic stem cells can give rise to all cell types, while adult stem cells are usually defined by a specific tissue which they can specify / progenitor cells divide from stem cells, and can divide into differentiated cells, but they cannot go "backwards" to stem cells / pluripotent stem cells can theoretically differentiate into any cell tissue / planaria seem to preserve pluripotency into adulthood after a certain number of radiations, few planaria could even live on to continue replicating cells / even a very small number of neoblasts can allow for regeneration in an organism

regulation of tissues is done hierarchically, done with the most broad-sense tissues down to more specific tissues

there are differences in what NC can "do" -- it can differentiate differently across different phylogenies (where the transition from trunk to jawed vertebrates started to become more defined)

do differences in the regionalization of the NC lead to different cell types arising?

inject a dye into a tissue that is still developing to see where that developing tissue ends up

trunk NC can give rise to mineralized tissues (why we find mineralized, fossilized shark teeth but no fossilized sharks, due to the jaws of sharks being developed from trunk NC)

beyond cartilaginous fish, the skeleton seems to come entirely from the mesoderm

So, does SS or NS act the most on this trait?

If swimming speed was also acted on by NS (not just SS), would it adhere to SK's definition?

Narrow-Sense Natural Selection "as long as you can stay in the game, you are succeeding at playing NS, but if you are winning the game, you are succeeding at playing SS"

If males are being chosen/directly competing against each other, the males are still being "chosen" because they are competing within the sex to get a mate or competing outside of their sex to be chosen

Darwin could not have been completely clear of how elaborate the diverse aspects of sexual selection there is/could be, he seemed hyper-focused on female choice/male competition